AUBF LECTURE

OUR LADY OF FATIMA UNIVERSITY

CSMLS

PRELIMS 2. Urine contains information, which can be obtained by

LESSON 1: HISTORY, inexpensive laboratory tests, about many of the body’s
QA/QC AND SAFETY IN major metabolic functions
THE CLINICAL LABORATORY
CLSI defines urinalysis as:
 “The testing of urine with procedures commonly performed
INTRODUCTION in an expeditious, reliable, accurate, safe, and cost-
 Analyzing urine was actually the beginning of laboratory effective manner.”
medicine.
Reasons for performing urinalysis identified by CLSI
PHYSICAL EVIDENCE include:
 Drawings of cavemen  aiding in the diagnosis of disease
 Egyptian hieroglyphics  screening asymptomatic populations for undetected
disorders
BASIC OBSERVATION:  monitoring the progress of disease and the effectiveness of
 Color therapy
 Turbidity
 Odor QUALITY CONTROL AND
 Volume QUALITY ASSESSMENT
 Viscosity DEFINITION
 Sweetness  Quality Assessment - refers to the overall process of
guaranteeing quality patient care and is regulated
INTRODUCTION throughout the total testing system.
 modern urinalysis has expanded beyond physical  Quality system - refers to all of the laboratory’s policies,
examination of urine include: processes, procedures, and resources needed to achieve
o chemical analysis quality testing.
o microscopic examination
Quality Assessment program includes:
HISTORY  Testing of controls
 5th century BC – Hippocrates wrote a book on “uroscopy.”  Pre-analytical factors
 1140 AD - color charts had been developed that described  Analytical factors
the significance of 20 different colors.  Post-analytical factors
 1627 - Thomas Bryant published a book about “pissed
prophets.” o QA is the continual monitoring of the entire test process
 1694 - Frederik Dekkers discover albuminuria. from test ordering and specimen collection through
 17th century - invention of the microscope reporting and interpreting results.
 1827- concept of urinalysis as part of a doctor’s routine
patient examination LIST OF LABORATORY ACCREDITATION
AGENCIES:
 Joint Commission on the Accreditation of Healthcare
Organizations
 College of American Pathologists
 American Association of Blood Banks
 American Osteopathic Association
 American Society of Histocompatibility and
Immunogenetics
 Commission on Laboratory Assessment

URINALYSIS PROCEDURE MANUAL

 A procedure manual containing all the procedures
performed in the urinalysis section must be available and
must comply with the CLSI guidelines.

The following information is included for each procedure

clinical significance
 patient preparation
 specimen type
 method of collection
 1930 - Urinalysis began to disappear from routine  specimen acceptability and criteria for rejection
examinations.  Reagents
 step-by-step procedure
Two unique characteristics of a urine specimen  recording of results
1. Urine is a readily available and easily collected specimen

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 PREANALYTICAL FACTORS  QC procedures are performed to ensure that acceptable
 Specific information on specimen collection and handling standards are met during the process of patient testing.
should be stated at the beginning of each procedure.  Control results must be recorded in a log, either paper or
electronic.
REQUISITION FORM CONTENT:
 Actual date and time TYPE OF QUALITY CONTROL
 Preservation technique EXTERNAL QUALITY CONTROL
 Time received and performed  Are used to verify the accuracy and precision of a test and
 Tests requested are exposed to the same conditions as the patient samples.
 Patient identification  External controls are tested and interpreted in the
laboratory by the same person performing the patient
testing.

INTERNAL QUALITY CONTROL

 Consists of internal monitoring systems built in to the test
system.

ANALYTICAL FACTORS
 The analytical factors are the processes that directly affect
the testing of specimens.
o It include:
 reagents Competency of Personnel and Facilities
 Instrumentation  Quality control is only as good as the personnel performing
 Procedure and monitoring it.
 QC  Personnel must understand the importance of QA.
 Competency of personnel performing the tests  Documentation of continuing education must be
maintained.
Reagents
 An adequate, uncluttered, safe working area is also
 All reagents and reagent strips must be properly labeled essential for both quality work and personnel morale.
with the date of preparation or opening, purchase and
received date, expiration date, and appropriate safety
POST-ANALYTICAL FACTORS
information.
 Postanalytical factors are processes that affect the
 Reagents are checked daily or when tests requiring their
reporting of results and correct interpretation of data.
use are requested
It include:
INSTRUMENTATION AND EQUIPMENT  Reporting of Results
 The most frequently encountered instruments: o Standardized reporting formats
o Refractometers o Electronic transmission is now the most common
o Osmometers method for reporting results.
o Automated reagent strip readers  Interpretation of Results
o Automated microscopy instruments
o All known interfering substances should be listed for
o Refrigerators
evaluation of patient test data.
o Centrifuges
o Microscopes
o Water baths.

Procedure
 concise testing instructions are written in a step-bystep
manner

Quality Control
 Refers to the materials, procedures, and techniques that
monitor the accuracy, precision, and reliability of a
laboratory test.

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 BIOLOGICAL HAZARDS
 The health-care setting provides abundant sources of
potentially harmful microorganisms.
 Understanding how microorganisms are transmitted is
essential in preventing infection.
 The CHAIN OF INFECTION is a continuous link between a
source, a method of transmission, and a susceptible host.

 Preventing completion of the chain of infection is a primary

objective of biological safety.
 Proper handwashing and wearing personal protective
equipment are important.
 In 1987 the CDC instituted Universal Precautions.

SAFETY IN THE CLINICAL

LABORATORY
INTRODUCTION
 The clinical laboratory contains a variety of safety hazards, UNIVERSAL PRECAUTIONS
many of which are capable of producing serious injury or
 Under UP all patients are considered to be possible carriers
life threatening disease.
of blood-borne pathogens.
 To work safely in the laboratory you must learn: o guideline recommends:
o hazards exist  wearing gloves in collection of specimen
o basic safety precautions  wearing face shields (mask)
o How to apply the basic rules of COMMON SENSE  Puncture resistant containers.
required for everyday safety.
BSI GUIDELINES
TYPES OF SAFETY HAZARDS
 Consider all body fluids and moist body substances to be
TYPE SOURCE POSSIBLE INJURY
potentially infectious.
Biological Infectious agents Bacterial, fungal,
o guideline recommends:
viral, or parasitic
 Wear gloves at all times
infections
 Do not require hand washing following removal of
Sharps Needles, lancets, Cuts, punctures,
gloves unless visual contamination is present.
broken glass or blood-borne
pathogen
BIOLOGICAL HAZARDS
exposure  In 1996 the CDC create the new guideline called
Chemical Preservatives and Exposure to toxic, STANDARD PRECAUTIONS.
reagents carcinogenic, or  Standard Precautions are as follows
caustic agents  Handwashing
 Gloves
Radioactive Equipment and Radiation
 Mask, eye protection, and face shield
radioisotopes exposure
 Laboratory Gown
Electrical Ungrounded or Burns or shock
 Patient care equipment
wet equipment;
 Environmental control
frayed cords
Linen
Fire/Explosive Bunsen burners, Burns or  Occupational health and blood-borne pathogens
organic chemicals dismemberment  Patient placement
Physical Wet floors, heavy Falls, sprains, or
boxes, patients strains
 The Occupational Exposure to Blood-Borne Pathogens
Standard is a law monitored and enforced by OSHA.

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 Specific requirements of this OSHA standard include the
following:
o 1. Requiring all employees to practice UP/Standard
Precautions
o 2. Providing PPE to employees
o 3. Providing sharps disposal containers and prohibiting
recapping of needles
o 4. Prohibiting eating, drinking, smoking, and applying
cosmetics, lip balm, and contact lens in the work area
o 5. Labeling all biohazardous material and containers
o 6. free immunization for HBV
o 7. daily disinfection protocol for work surfaces
o 8. Providing medical follow-up for employees who have
been accidentally exposed to blood-borne pathogens
o 9. Documenting regular training in safety standards for
employees.
PERSONAL PROTECTIVE EQUIPMENT
 PPE includes:
o gloves
o fluid-resistant gowns
o goggles and mask
HANDWASHING
 Hand contact is the primary method of infection
transmission.
 Correct hand washing technique includes the following
steps:
o 1. Wet hands with water
o 2. Apply soap
o 3. Rub to form a lather, create friction, and loosen
debris.
o 4. Clean between fingers, including thumbs, under
fingernails and rings, and up to the
o wrist, for at least 15 seconds.
o 5. Rinse hands in a downward position
o 6. Dry with a paper towel
o 7. Turn off faucets with a clean paper towel
DISPOSAL OF BIOLOGICAL WASTE
 All biological waste, except urine, must be placed in
appropriate containers labeled with the biohazard symbol.
 The waste is then decontaminated following institutional
policy.
o (Example. incineration, autoclaving etc…)
 Urine may be discarded by pouring it into a laboratory sink.
 Disinfection of the sink using sodium hypochlorite should be
performed daily.
SHARP HAZARDS
 All sharp objects must be disposed in puncture resistant
containers.
 Puncture-resistant containers should be conveniently
located within the work area
CHEMICAL HAZARDS
 Every chemical in the workplace should be presumed
hazardous.
CHEMICAL SPILLS
 flush the area with large amounts of water for at least 15
minutes and seek medical attention
 Contaminated clothing should be removed as soon as
possible.
 Chemical spill kits should be always available for cleaning
up spills.
CHEMICAL HANDLING
 Chemicals should never be mixed together unless specific
instructions are followed and they must be added in the
order specified.
CHEMICAL HYGIENE PLAN
 Required by OSHA
 Purpose of the plan:
o 1. Appropriate work practices
o 2. Standard operating procedures
o 3. PPE
o 4. Engineering controls (fume hoods, safety cabinets,
etc…)
o 5. Employee training requirements
o 6. Medical consultation guidelines
CHEMICAL LABELING
o Hazardous chemicals should be labeled with a
description of their particular hazard.
o The NFPA has developed the Standard System for the
Identification of the Fire Hazards of Materials
o This symbol system is used to inform fire fighters of the
hazards they may encounter with fires in a particular
area.
MATERIAL SAFETY DATA SHEETS
 The OSHA Federal Hazard Communication Standard
requires that all employees have a right to know about all
chemical hazards present in their workplace.
 Information contained in an MSDS includes the following:
o 1. Physical and chemical characteristics
o 2. Fire and explosion potential
o 3. Reactivity potential
o 4. Health hazards and emergency first aid procedures
o 5. Methods for safe handling and disposal
RADIOACTIVE HAZARDS
 Radioactivity is encountered when procedures using
radioisotopes are performed.
 The amount of radiation exposure is related to a
combination of time, distance, and shielding.
 Exposure to radiation during pregnancy presents a danger
to the fetus
PRECAUTIONS:
 Equipment should not be operated with wet hands.
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 Equipment that has become wet should be unplugged and
allowed to dry completely before reusing.
 Equipment also should be unplugged before cleaning.

 What to do when electrical shock occurs?

o electrical source must be removed immediately
o Turning of the circuit breaker
o Unplugging the equipment\
o Use of nonconductive material to move the equipment.

FIRE/EXPLOSIVE HAZARDS
 JCAHO requires that all health-care institutions post
evacuation routes and detailed plans to follow in the event
of a fire.
o Rescue
o Alarm
o Contain
o Extinguish

PRECAUTIONS:
 Flammable chemicals should be stored in safety cabinets
 Explosion-proof refrigerators and cylinders of compressed
gas should be located away from heat and securely
fastened to prevent accidental capsizing.
 Fire blankets should be present in the laboratory.

The NFPA classifies fires with regard to the type of burning

material.

FIRE EXTINGUISHING TYPE OF EXTINGUISHER

TYPE MATERIAL FIRE/COMPOSITION
OF FIRE
CLASS A Wood, paper, Class A Water
clothing
CLASS B Flammable Class B Dry chemicals,
organic carbon
chemicals dioxide, foam,
or halon
CLASS C Electrical Class C Dry chemicals,
carbon
dioxide, or
halon
CLASS D Combustible None Sand or dry
metals powder
Class ABC Dry chemicals

 It is important to be able to operate the fire extinguishers.

REMEMBER PASS
o Pull
o Aim
o Squeeze
o Sweep

GENERAL PRECAUTIONS:
 Avoid running in rooms and hallways
 Watch for wet floors
 Bend the knees when lifting heavy objects
 Keep long hair pulled back
 Avoid dangling jewelry
 Maintain a clean and organized work area.
 Wear closed-toe shoes

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AUBF LECTURE
OLFU PAMPANGA

RENAL FUNCTION AND INTRODUCTION TO

URINALYSIS
HISTORY
 Basic observations
 Color
 Turbidity
 Odor THOMAS ADDIS
 Volume  Methods for quantitating the microscopic sediment.
 Viscosity
 Sweetness (by noting that certain specimens attracted ants RICHARD BRIGHT
or tasted sweet).
 Introduced the concept of urinalysis as part of a doctor’s
routine patient examination in 1827.

URINE
2 UNIQUE CHARACTERISTICS OF URINE
SPECIMEN:
 1. Urine is a readily available and easily collected specimen.
 2. Urine contains information, which can be obtained by
inexpensive laboratory tests, about many of the body’s
major metabolic functions.

FACTORS THAT AFFECT URINE CONCENTRATION

 Dietary intake
 Physical activity,
 Body metabolism,
HIPPOCRATES
 Endocrine functions.
 Father of medicine
 Wrote a book on uroscopy in 5th BCE
URINE COMPOSITION
o Physicians concentrated their efforts very intensively  95% water
on the art of uroscopy, receiving instruction in urine  5% solutes
examination as part of their training.

FREDERIK DEKKERS
 Discovery in 1694 of albuminuria by boiling urine. UREA
 Produced in the liver
 Accounts nearly half amount of dissolved solids in the urine.

CHLORIDE
 Major inorganic substance in the urine

SODIUM AND POTASSIUM

 2nd to chloride

o High urea and creatinine content can identify a fluid as

urine.
THOMAS BRYANT (1627)
 “ Pisse Prophets ” URINE VOLUME
 The revelations in this book inspired the passing of the first  Normal daily urine output is usually 1200 to 1500 ML.
medical licensure laws in England  A range of 600 to 2000 mL is considered normal.

ANTON VAN LEEUWENHOEK ABNORMAL URINE VOLUME

 17th century MICROSCOPE OLIGURIA

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 DECREASE IN URINE OUTPUT  Begin and end with an empty bladder
 < 1ML/KG/HR : INFANTS
 <0.5ML/KG/HR : CHILDREN 3 GLASS COLLECTION METHOD
 <400ML/DAY : ADULTS  1st glass – first urine
 2nd glass – midstream
ANURIA  3rd glass – prostatic massage
 Cessation of urine  Result 3rd glass will have a result of wbc/hpf 10 times
 Serious damage to the kidneys or higher
 Decrease in the flow of blood to the kidneys.  2nd glass. Kidney infection

POLYURIA PPMT
 increase in  1st sample midstream clean catch
 daily urine volume  2nd sample prostate is massage
 > 2.5L/day : ADULTS  Bacteria reading 10times higher than the first is considered
 > 2-3mL/kg/day : Children prostatic infection

o The kidneys excrete two to three times more urine DRUG SPECIMEN COLLECTION
during the day than during the night. An increase in the  Chain of custody
nocturnal excretion of urine is termed nocturia.  30 to 45mL of urine is collected
 Urine temp must be taken within 4 mins 32.5-37.7
SPECIMEN COLLECTION
 Urine is a biohazardous substance that requires the RENAL FUNCTION
observance of Standard Precautions. INTRODUCTION
 Specimens must be collected in clean, dry, leak-proof
containers. 50mL capacity URINARY SYSTEM
 Properly applied screw-top lids are less likely to leak than KIDNEYS
are snap-on lids.  Bean-shaped organ located in the posterior wall of the
 Patient’s name and identification number, the date and time abdomen. (retroperitoneum)
of collection, and additional information such as the
patient’s age and location and the healthcare provider’s URETERS
name
 25 cm long , carry the urine from the kidney to the bladder
 Unacceptable situations include:
URINARY BLADDER
o 1. Specimens in unlabeled containers
o 2. Nonmatching labels and requisition forms  shaped like a 3 sided pyramid, stores the urine produced
o 3. Specimens contaminated with feces or toilet
paper URETHRA
o 4. Containers with contaminated exteriors  4cm long in women
o 5. Specimens of insufficient quantity  24cm long in men
o 6. Specimens that have been improperly  Deliver the urine for excretion.
transported
o Laboratories should have a written policy detailing KIDNEY
their conditions for specimen rejection.  Primary function
1. clear waste products
SPECIMEN INTEGRITY 2. reabsorption of nutrients
o Contains approximately 1 to 1.5 million nephrons.

 Kidney is about
o 12.5cm in length
o 6cm in width
o 2.5cm in depth

CORTICAL NEPHRONS
 1. make up approximately 85% of nephrons
 2. primarily in the cortex of the kidney
 3. for removal of waste products and reabsorption of
nutrients.

SPECIMEN TYPES JUXTAMEDULLARY NEPHRONS

RANDOM SPECIMEN  1. have longer loop of henle
 Most commonly received specimen  2. concentration of urine

24 HR. RENAL FUNCTION

 For detection of solutes with diurnal variation  Renal blood flow
 Ex. – Catecholamines  Glomerular filtration
 17-hydroxysteroids lowest early morning highest afternoon  Tubular reabsorption
electrolytes  Tubular secretion

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 RENAL BLOOD FLOW
 The renal artery supplies blood to the kidney.
 Approximately 25% of the blood from the heart goes to the
kidney.
 Blood enters the capillaries of the nephron through the
afferent arteriole and exit in efferent arteriole
 total renal blood flow = 1200 mL/min
 total renal plasma flow =600 to 700 mL/min
 RENAL artery → afferent arteriole → glomerulus → efferent
arteriole → peritubular capillaries → vasa recta → renal
vein
GLOMERULAR FILTRATION
GLOMERULUS
 Consists of a coil of approximately eight capillary lobes.
 Located within Bowman’s capsule.
 nonselective filter of plasma substances <70,000 MW
CAPILLARY TUFT
 collectively term for eight capillary lobes
BOWMANS CAPSULE
 Beginning of the renal tubule.
o The varying sizes of these arterioles help to create the
hydrostatic pressure differential important for
glomerular filtration and to maintain consistency of
glomerular capillary pressure and renal blood flow
within the glomerulus.
GLOMERULAR FILTRATION
 Filtration process include:
o Cellular Structure of the Glomerulus
o hydrostatic and oncotic pressures
o Renin-angiotensin-aldosterone system
 Plasma filtrate must pass through three cellular layers:
1. capillary wall membrane
2. basement membrane (basal lamina)
3. visceral epithelium of Bowman’s capsule (
podocytes )
 The endothelial cells of the capillary wall differ from those
in other capillaries by containing pores and are referred to
as fenestrated.
 Podocytes are the intertwining foot processes of the inner
layer of Bowman’s capsule.
 The barrier contains a shield of negativity that repels
molecules with a positive charge.
CELLULAR STRUCTURE OF THE GLOMERULUS
HYDROSTATIC AND ONCOTIC PRESSURES
 Presence of hydrostatic pressure enhances the filtration.
This pressure is necessary to overcome the opposition of
pressures from the fluid within Bowman’s capsule and the
oncotic pressure of unfiltered plasma proteins in the
glomerular capillaries.
 Dilation of the afferent arterioles and constriction of the
efferent arterioles when blood pressure drops prevent a
marked decrease in blood flowing through the kidney.
 Increase in blood pressure results in constriction of the
afferent arterioles to prevent over filtration or damage to the
glomerulus.
RENIN ANGIOTENSIN ALDOSTERONE SYSTEM
RAAS regulates the flow of blood to and within the
glomerulus.
 The system responds to changes in blood pressure and
plasma sodium content that are monitored by the
juxtaglomerular apparatus.
 Juxtaglomerular apparatus consist of:
o Juxtaglomerular cells ( afferent arteriole)
o Macula densa ( distal tubule )
 Low plasma sodium content decreases water retention
within the circulatory system, resulting in a decreased
overall blood volume and subsequent decrease in blood
pressure.
 RENIN → reacts w/ blood borne SUBSTRATE
ANGIOTENSINOGEN → ANGIOTENSIN I → circulates →
ALVEOLI of the lungs contain ACE → (active form)
ANGEOTENSIN II corrects renal blood flow in the ff ways:
vasodilation of the afferent arterioles and constriction of the
efferent arterioles stimulating reabsorption of sodium and
water in the proximal convoluted tubules trigger the release
of sodium retaining hormone ALDOSTERONE produce by
adrenal cortex ADH by the hypothalamus.
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 Must to know about glomerular filtration:
o 1. Every minute approximately two to three million
glomeruli filter passes the glomerulus.
o 2. This filtration is nonselective and can be
differentiated to plasma by the absence of the plasma
proteins
o 3. Analysis of the fluid as it leaves the glomerulus
shows the filtrate to have a specific gravity of 1.010 and
confirms as an ultrafiltrate of plasma.
TUBULAR REABSORPTION
 The body cannot lose 120 mL of water-containing essential
substances every minute.
 Proximal convoluted tubule is the one responsible of
reabsorbing most of the essential substances and water
needed by the body.
 Active transport, like passive transport, can be influenced
by the concentration of the substance being transported.
 (Tm) - maximal reabsorptive capacity
 The plasma concentration at which active transport stops
is termed the renal threshold.
 Knowledge of the renal threshold and the plasma
concentration can be used to distinguish between excess
solute filtration and renal tubular damage.
TUBULAR CONCENTRATION
 DLH and ALH IS THE BEGINNING OF RENAL
CONCENTRATION because the filtrate is exposed to the
high osmotic gradient of the renal medulla.
 Water is removed by osmosis in the descending loop of
Henle.
 Sodium and chloride are reabsorbed in the ascending loop.
OSMOSIS
 A process by which molecules of solvent tend to pass
through a semipermeable membrane from a less
concentrated solution into a more concentrated one.

 The final concentration of the filtrate begins in the late distal

convoluted tubule and continues in the collecting duct.
 Reabsorption depends on the osmotic gradient in the
medulla and the hormone vasopressin.
 Production of ADH is determined by the state of body
hydration.
 ADH renders the walls of the distal convoluted tubule and
collecting duct permeable or impermeable to water.

TUBULAR REABSORPTION
 Reabsorption Mechanisms
 Tubular Concentration
 Collecting Duct Concentration

REABSORPTION MECHANISMS
ACTIVE TRANSPORT LOW TO HIGH
 Need to be combined with a carrier protein contained in the
membrane of the renal tubule epithelial cells.
 Glucose, Amino acid, salts, chloride, and sodium.
 GAAS – PCT
 CHLO- ALH
TUBULAR SECRETION
 SOD- PCT AND DCT
 Involves the passage of substances from the blood in the
peritubular capillaries to the tubular filtrate.
PASSIVE TRANSPORT HIGH TO LOW
 2 major functions:
 Movement of molecules across a membrane as a result of
o Eliminating waste products not filtered by the
differences in their concentration or electrical potential on
glomerulus.
opposite sides of the membrane.
o Regulate acid–base balance.
 WATER, UREA, AND SODIUM
 WATER- PCT DLH CD  Medications, cannot be filtered by the glomerulus because
 UREA- PCT ALH they are bound to plasma proteins.
 SODIUM- ALH  The major site for removal of these nonfiltered substances
is the PCT.

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  Standard test used to measure the filtering capacity of the
glomeruli.
 Measures the rate at which the kidneys are able to remove
a filterable substance from the blood.
 Characteristics of substance to be analyzed
o Substance is neither reabsorbed nor secreted by the
tubules.
o Stability in urine for 24hrs of collection
o Plasma level consistency
o Availability to the body
o Availability of test for analysis of the substance.
 Ex. UREA, CREATININE, INULIN, BETA2
ACID BASE BALANCE MICROGLOBULIN, CYSTATIN C AND
 Important to maintain blood pH. RADIOISOTOPES.
 Blood must buffer and eliminate the excess acid formed by
dietary intake and body metabolism. UREA
 The buffering capacity of the blood depends on  READILY AVAILABLE IN ALL URINE
bicarbonate.  METHODS ARE READILY AVAILABLE
 100% reabsorption of filtered bicarbonate and occurs  40% ARE BEING REABSORBED
primarily in the proximal convoluted tubule.
 Hydrogen ions are readily filtered and reabsorbed. INULIN CLEARANCE
 Normal blood pH =7.4  A polymer of fructose.
HCO3  stable substance
 filtered by the glomerulus  not a normal body constituent
 returned to blood to maintain blood ph  Reference method for clearance test.
 H2CO3 – carbonic acid
CREATININE CLEARANCE
 routinely used
 A waste product of muscle metabolism that is normally
found at a relatively constant level in the blood.

INJECTION OF RADIONUCLEOTIDES
 Method of determining glomerular filtration through the
plasma disappearance of the radioactive material and
enables visualization of the filtration in one or both kidneys

BETA-2-MICROGLOBULIN
 Dissociates from human leukocyte antigen at a constant
RENAL FUNCTION TESTS rate and is rapidly removed from the plasma by glomerular
 Glomerular Filtration Tests filtration.
 Tubular Reabsorption Tests  MW= 11,800
 Tubular Secretion and Renal Blood Flow Tests  ENZYME IMMUNOASSAY is use for measurement.
 inc. plasma level= dec. GFR
GLOMERULAR FILTRATION RATE/TEST  Not reliable for patient w/ immunologic disorders or
 Clearance test ( inulin and creatinine ) malignancy

TUBULAR REABSORPTION TEST CYSTATIN C

 Free water clearance test  Produced by all nucleated cells
 Osmolarity ( freezing point osmometer and vapor pressure  Filtered and reabsorbed and broken down by the renal
osmometer ) tubular cells.
 No cystatin C is secreted and serum concentration can be
TUBULAR SECRETION AND RENAL BLOOD FLOW directly related to GFR.
TEST  Immunoassay are use for measuring cystatin C
 P-amminohippuric acid test  Recommended for pediatric , diabetic, elderly and critically
 Titratable acidity and urinary ammonia. ill patients.

GLOMERULAR FILTRATION RATE

 Greatest source of error is the use of improperly timed urine
specimens.
 The GFR is reported in milliliters cleared per minute
 To calculate you need:
 Urine Volume in ml/min
 Urine creatinine concentration in mg/dl
 Plasma creatinine concentration in mg/dl
GLOMERULAR FILTRATION TESTS  Normal creatinine clearance
CLEARANCE TEST  men: 107-139 ml/min women: 87-107 ml/min

10
 Normal plasma creatinine range: 0.5-1.5mg/dl
GLOMERULAR FILTRATION TESTS
CREATININE CLEARANCE DISADVANTAGES
 Can be secreted by the tubules
 Secretion is affected by medication
 Bacteria will break down urinary creatinine
 Dietary
 Cannot be used in patient with muscle-wasting disease /
athletes
 It needs correction for body surface area especially in
children.
o Newer methods that do not require the collection of
timed (24-hour) using serum creatinine, cystatin c or
beta2-macroglobulin values.
MEDICATIONS
 Gentamicin
 Cephalosporin
 Cimetidine ( Tagamet) inhibit tubular secretion of creatinine
 estimated glomerular filtration rate (eGFR)
GLOMERULAR FILTRATION RATE
CLINICAL SIGNIFICANCE OF CREATININE
CLEARANCE
 Used to assess the functioning nephrons but also but also
the functional capacity of these nephron.
 Used to determine the extent of nephron damage in known
cases of renal disease.
 Monitor the effectiveness of the treatment
 Feasibility of administering medications.
2. PHENOLSULFONPHTHALEIN
 NOT CURRENTLY PERFORMED
 STANDARDIZATION AND INTERPRETATION OF
RESULT IS DIFFICULT
 COLORLESS IN ACID SOLUTION AND RED IN
ALKALINE SOLUTION
3. URINARY AMMONIA AND TITRATABLE ACIDITY
 DETERMINES THE DEFECTIVE FUNCTION IN THE
ABILITY OF THE KIDNEY TO PRODUCE ACIDIC URINE.
 FACTORS INVOLVE TO PRODUCE AN ACIDIC URINE
A. TUBULAR SECRETION OF HYDROGEN ION
B. DCT PRODUCTION AND SECRETION OF AMMONIUM
IONS
 A NORMAL PERSON EXCRETES 70meq/day of acid in
the form of either titratable acid, hydrogen phosphate ions ,
ammonium ions.
 A diurnal variation affects urine acidity
 Renal tubular acidosis is the inability to produce an acidic
urine in the presence of metabolic acidosis.
 Specimen: fresh or toluene preserved urine.
 Cause by
o Impaired tubular secretion PCT
o Impaired ammonia secretion DCT

TUBULAR REABSORPTION TEST

TEST:
FISHBERG CONCENTRATION TEST
 Deprived of fluids for 24 hrs prior to measuring SG.

MOSENTHAL CONCENTRATION TEST

 Compared volume and SG of day and night urine sample

OSMOMETRY
 Test use for quantitative measurement of renal
concentration ability.
 Reported in milliosmole (mOsm)

TUBULAR REABSORPTION TEST

 Loss of reabsorption capability is often the first function
affected in renal disease.
 A.k.a CONCENTRATION test
 Urine concentration is largely determined by the body’s
state of hydration.

TUBULAR SECRETION AND RENAL BLOOD FLOW

TEST
TEST:
1. P-AMINOHIPPURIC ACID TEST
 EXOGENOUS PROCEDURE
 IS A NON TOXIC SUBSTANCE LOOSELY BOUND TO
PLASMA PROTEINS
 ALL PLASMA PAH IS SECRETED BY THE PCT,
THEREFORE THE VOLUME OF PLASMA DETERMINES
THE AMOUNT OF PAH OF THE URINE.

11
AUBF LECTURE YELLOW ORANGE
OLFU PAMPANGA  Bilirubin
o yellow foam appears when the specimen is shaken.
o Urobilinogen
PHYSICAL EXAMINATION
o it is due to photo-oxidation of urobilinogen to urobilin.
IMPORTANCE OF PHYSICAL EXAMINATION  Phenazopyridine (Pyridium)
 It provides a preliminary information concerning disorders. o drug use for UTI
 It aids in the evaluation of renal tubular function. o thick, orange pigment that interferes in chemical tests
 It is use to confirm or explain clinical findings in the chemical that are based in color reactions.
and microscopic analysis.
YELLOW GREEN
DISORDERS  due to photo-oxidation of bilirubin to biliverdin.
 Glomerular bleeding
 Liver disease BROWN / BLACK
 Inborn error of metabolism  RBC remaining in an acidic urine produce a brown color
 UTI due to oxidation of hemoglobin to methemoglobin.
 Glomerular bleeding
COLOR  Melanin
NORMAL URINE COLOR  Homogentinsic Acid
 Variation of urine color may be due to:  Other causes: levodopa, methyldopa, phenol derivatives &
 Normal metabolic functions metronidazole (Flagyl).
 Physical activities
 Dietary intake  Brown – oxidation of hemoglobin to methemoglobin.
 Pathologic disorders  Fresh brown urine containing blood – glomerular
 UROCHROME bleeding.
 Responsible for yellow color of urine.  Melanin – oxidation product of the colorless pigment
 Product of endogenous metabolism and is dependent in melanogen, produced in excess when a malignant
body̕ s metabolic state. melanoma is present.
 Homogentinsic ACID – metabolite of phenylalanine , black
SIDE NOTES: color in alkaline urine from person with IEM called
 Thudichum named urochrome in 1864 alkaptonuria.
 urochrome is dependent on the body’s metabolic
state, with increased amounts produced in thyroid BLUE
conditions and fasting.  Medications like methocarbamol, methylene blue, and
 Urochrome also increases in urine that stands at amitriptyline
room temperature.
GREEN
 Other pigments responsible for the color of normal urine  clorets
are:  phenol derivatives found in IV medications
1. Uroerythrin
 Pseudomonas species
2. Urobilin

SIDE NOTES:
PURPLE
 Uroerythrin  indicanuria
- Pink pigment  bacterial infection
- Indicator that the specimen was refrigerated.
 Urobilin  Methocarbamol (robaxin) – muscle relaxant
- Oxidation product of urobilinogen  Amitriptyline ( Elavil ) – antidepressant
- Imparts Orange brown color of urine that is not fresh  Methylene blue – fistulas
 purple staining may occur in catheter bags and is caused
ABNORMAL URINE COLOR by
DARK YELLOW  indican in the urine or a bacterial infection, frequently
 concentrated specimen caused
 by Klebsiella or Providencia species
AMBER
 dehydration from fever and burns RED
 Can be due to RBC, hemoglobin, myoglobin, menstrual
o Normal urine produce small amount of foam and contamination, rifampin, phenolphthalein, phenindione,
disappear rapidly phenothiazines, beets and blackberries.
o Presence of large amount of protein produce white
foam o Rbc- Cloudy urine , positive in chemical test for blood,
 Bilirubin – can be detected in chemical analysis rbc observed microscopically
 Urobilinogen – no yellow foam is seen when shaken. o Hgb - clear urine w positive chem test , intravascular
 Pyridium- produce yellow foam when shaken. Mistaken as hemolysis
o Mgb- clear urine w/ positive chemical test , muscle
bilirubin
damage
o Beets- red in alkaline urine
o Blackberries - red in acidic urine

12
 PORT WINE SPECIFIC GRAVITY
 Due to oxidation of porphobilinogen to porphyrins.  Instruments use :
o Urinometer
CLARITY o Harmonic Oscillation Densitometry (HOD)
 Clarity is the general term that refers to o Refractometer
transparency/turbidity of a urine. o Chemical reagent strip
 Precipitation of amorphous phosphates and carbonates
may cause white cloudiness.  Urinometer and HOD are direct method in determining the
 Common terminology use: Specific Gravity.
o Clear - no visible particulates, transparent  Refractometer and Chemical rgt strip are the indirect
o Hazy - few particulates , print easily seen MTD.
o Cloudy - many particulates , print blurred
o Turbid - print cannot be seen through urine URINOMETER
o Milky - may precipitate or be clotted  Consist of a weighted float that displaces a volume of liquid
equal to its weight.
PATHOLOGIC VS.NON-PATHOLOGIC TURBIDITY  Disadvantages :
 Causes: o Less accurate
o Large volume needed - 10-15mL
 Non  Pathologic o Temperature correction needed
Pathologic
 Urinometer ( Hydrometer )
 Squamous  Rbc , WBC  CLSI – clinical and laboratory standards institute formerly
epithelial cells and
NCCLS
Bacteria
 National committee for clinical laboratory standards
 Mucus  Yeast ,
abnormal REFRACTOMETER
crystals  Principle use is refractive index.
 Advantages:
 Amorphous  Lymph fluid o small volume of urine needed
phosphate o no temperature corrections
 Calibrators:
 Amorphous  lipids o distilled water
carbonates o 5% NaCl
o 9% sucrose
 Amorphous 
urates
HARMONIC OSCILLATION DENSITOMETRY
 semen , fecal   It is based on the principle that the frequency of sound wave
contamination, entering a solution changes in proportion to the density of
radio graphic the solution.
contrast
media,

 talcum 
powders and
vaginal smear

 Amorphous phosphate and carbonates – white ppt in

alkaline pH
 Amorphous urates – resembles as pink brick dust due to
uroerythyrin in acidic pH
 Rbc , wbc and bacteria – cause by infection or a systemic
organ disorder
CHEMICAL REAGENT STRIP
 Principle is based on the pKa changes of a polyelectrolyte.
 pKa ( dissociation constant)

CLINICAL SIGNIFICANCE
 The specific gravity entering the glomerulus is 1.010.
 Isosthenuric - 1.010
 Hyposthenuric - <1.010
 Hypersthenuric - > 1.010
 Normal random urine SG range is 1.003-1.035
 Below 1.003 is not a urine
 Above 1.035 seen in * IV pyelogram, dextran

13
 ODOR
 It is not part of routine urinalysis.
Odor Cause

 Aromatic  normal

 Foul –  Uti , bacterial decomposition

ammonia-
like

 Fruity,  DM, starvation , vomiting

sweet

 Maple  Maple syrup urine disease

syrup

 Mousy  Phenylketonuria

 Rancid  Tyrosinemia

 Sweaty  Isovaleric acidemia

feet

 Cabbage  Methionine malabsorption

 Bleach  Contamination

14
AUBF LECTURE
OLFU PAMPANGA
CHEMICAL EXAMINATION: PART 1
REAGENT STRIP
 Provide a simple, rapid means for performing medically
significant chemical analysis of urine.
 Glucose , Bilirubin , Ketones, Specific Gravity, Protein , pH
, Blood , Urobilinogen , Nitrite, Leukocyte esterase
 Major Company that produce commercially available
reagent strips:
o Siemens Medical Solutions Diagnostics –
tradename multistix
o Roche Diagnostics – chemstrip
o Reagent strips consist of chemical-impregnated
absorbent pads attached to a plastic strip.
o A color-producing chemical reaction takes place when
the absorbent pad comes in contact with urine
REAGENT STRIP TECHNIQUE
 Mix → Dip → Remove → Blot → Compare
IMPROPER TECHNIQUES:
 Unmixed specimens - Formed elements such as red and
white blood cells sink to the bottom of the specimen and will
be undetected
 Allowing the strip to remain in the urine for extended period
- leaching of reagents from the pads
 Excess urine remaining on the strip - run-over between
chemicals on adjacent pads, producing distortion of the
colors – blot the edge to prevent
 Timing
 Not having a good light source
 Not holding the strip close enough to the color chart without
actually being placed on the chart.
 Reagent strips and color charts from different
manufacturers are not interchangeable
 Specimens that have been refrigerated must be allowed to
return to room temperature - enzymatic reactions on the
strips are temperature dependent.
HANDLING AND STORAGE
 Store w/ desiccant in an opaque, tightly closed container
 Remove strip prior to use and reseal immediately
 Stored below 30˚c / room temperature
 Bottle should not be open in the presence of volatile fumes.
 Do not use expired reagent strips.
 Care must be taken not to touch the chemical pads
CAUSES OF SPECIMEN DETERIORATION:
 MOISTURE
 VOLATILE CHEMICALS
 HEAT
 LIGHT
QUALITY CONTROL OF REAGENT STRIP
 EVERY 24 HRS
 Test open bottle of reagent strip w/ known positive and
negative controls every 24 hrs / beginning of each shift
 Resolve control results that are out of range by further
testing.
 Perform positive and negative controls on new reagents
and newly opened bottles of reagent strips.
 Record all control results and reagent lot numbers.
 Distilled water is not recommended as a negative control
because reagent strip chemical reactions are designed to
perform at ionic concentrations similar to urine.
CONFIRMATORY TESTING
 Confirmatory tests are defined as test using different
reagents or methodologies to detect the same substances
as detected by the reagent strips with the same or greater
sensitivity or specificity.
 Nonreagent strip testing procedures using tablets and liquid
chemicals – when questionable results are obtained or
highly pigmented specimens are encountered
REAGENT STRIP REACTION AND PRINCIPLE
PH
 Principle : Double Indicator SYSTEM
 Chemical Reaction :
CLINICAL SIGNIFICANCE
 aid in determining the existence of systemic acid–base
disorders of metabolic or respiratory origin
 management of urinary conditions that require the urine to
be maintained at a specific pH
 Identification of crystals observed during microscopic
examination of the urine sediment.
 The Multistix and Chemstrip brands of reagent strips
measure urine pH in 0.5- or 1-unit increments between pH
5 and 9.
 Methyl red – red to yellow in the pH range 4 to 6
 Bromthymol blue - yellow to blue in the range of 6 to 9
o **Therefore, in the pH range 5 to 9 measured by the
reagent strips, one sees colors progressing from
orange at pH 5 through yellow and green to a final deep
blue at pH 9
o **No known substances interfere with urinary pH
measurements performed by reagent strips.
15
 PROTEIN
 Principle : Protein error of indicator
 Chemical reaction:
 Most indicative of renal disease - require additional testing
to determine whether the protein represents a normal or a
pathologic condition
 Clinical proteinuria is indicated at 30 mg/dL or greater (300
mg/L).
SIDE NOTES
 Majority – albumin
 Others - Tamm-Horsfall protein (uromodulin) - renal tubular
epithelial cells; and proteins from prostatic, seminal, and
vaginal secretions
 **Contrary to the general belief that indicators produce
specific colors in response to particular pH levels,
 certain indicators change color in the presence of protein
even though the pH of the medium remains constant.
 This is because protein (primarily albumin) accepts
hydrogen ions from the indicator. The test is more sensitive
to albumin because albumin contains more amino groups
to accept the hydrogen ions than other proteins.
 Strip contains either tetrabromophenol blue (Multistix) or
3',3",5',5"-tetrachlorophenol, 3,4,5,6-
tetrabromosulfonphthalein (Chemstrip), and an acid buffer
to maintain the pH at a constant level.
 Both indicators appear yellow in the absence of protein;
however, as the protein concentration increases, the color
progresses through various shades of green and finally to
blue.
 Readings are reported in terms of negative, trace, 1+, 2+,
3+, and 4+;
o Highly buffered alkaline urine that overrides the acid
buffer system, False-positive readings are obtained
when the reaction does not take place under acidic
conditions.
GLUCOSE
 Principle: Double sequential enzyme REACTION
 Chemical Reaction:
GLYCOSURIA
 Glucose in the urine
 Under normal circumstances, almost all the glucose filtered
by the glomerulus is actively reabsorbed in the proximal
convoluted tubule; therefore, urine contains only minute
amounts of glucose - detection of hyperglycemia
OTHER NOTES
 impregnating the testing area with a mixture of glucose
oxidase, peroxidase, Chromogen, Buffer
 In the first step, glucose oxidase catalyzes a reaction
between glucose and room air (oxygen) to produce gluconic
acid and peroxide. In the second step, peroxidase catalyzes
the reaction between peroxide and chromogen to form an
oxidized colored compound that is produced in direct
proportion to the concentration of glucose.
 Reagent strip manufacturers use several different
chromogens, including potassium iodide (green to brown)
(Multistix) and tetramethylbenzidine (yellow to green)
(Chemstrip).
o ascorbic acid, that prevent oxidation of the chromogen
o High specific gravity and low temperature may
decrease the sensitivity of the test
KETONES
 Principle: Sodium nitroprusside Reaction
 Chemical Reaction:
 Ketonuria
o Ketones in the urine
 Represents three intermediate products of fat
metabolism:
o Acetone
o Acetoacetic acid
o B-hydroxybutyrate
 When the use of available carbohydrate as the major
source of energy becomes compromised, body stores of fat
must be metabolized to supply energy. Ketones are then
detected in urine.
 Clinical reasons for increased fat metabolism include the
inability to metabolize carbohydrate, as occurs in diabetes
mellitus
 Other name of principle: nitroferricyanide
 In this reaction, acetoacetic acid in an alkaline medium
reacts with sodium nitroprusside to produce a purple color.
 The test does not measure b -hydroxybutyrate and is only
slightly sensitive to acetone when glycine is also present
 Results are reported qualitatively as negative, trace, small
(1+), moderate (2+), or large (3+), or semiquantitatively as
negative, trace (5 mg/dL), small (15 mg/dL), moderate (40
mg/dL), or large (80 to 160 mg/dL).
16
  Bilirubin combine with 2,4-dichloroaniline diazonium salt or
2,6-dichlorobenzene-diazonium-tetrafluoroborate in an acid
medium to produce an azodye.
 Color: tan → pink → Violet
 Qualitative results are reported as negative, small,
moderate, or large, or as negative, 1+, 2+, or 3+.

BLOOD
 Principle: Pseudoperoxidase activity of hemoglobin
 Chemical Reaction:

o Bilirubin is an unstable compound that is rapidly photo-

oxidized to biliverdin when exposed to light. Biliverdin
 indicates the presence of red blood cells, hemoglobin, or does not react with diazo tests.
myoglobin o Nitrite may lower the sensitivity of the test, because
 pseudoperoxidase activity of hemoglobin to catalyze a they combine with the diazonium salt and prevent its
reaction between the heme component of both hemoglobin reaction with bilirubin.
and myoglobin and the chromogen tetramethylbenzidine to
produce an oxidized chromogen, which has a green-blue UROBILINOGEN
color.  Principle : Ehrlich’s Aldehyde Reaction (Multistix) and Azo-
 Reagent strip manufacturers incorporate peroxide, and coupling reaction (chemstrip)
tetramethylbenzidine, into the blood testing area. Two color  Chemical reaction:
charts are provided that correspond to the reactions that
occur with hemoglobinuria, myoglobinuria, and hematuria
 In the presence of free hemoglobin/myoglobin, uniform
color ranging from a negative yellow through green to a
strongly positive green-blue appears on the pad.
 In contrast, intact red blood cells are lysed when they come
in contact with the pad, and the liberated hemoglobin
produces an isolated reaction that results in a speckled
pattern on the pad.

 Seen in liver disease and hemolytic disorders.

Measurement of urine urobilinogen can be valuable in the
detection of early liver disease
 Multistix uses Ehrlich’s aldehyde reaction, in which
urobilinogen reacts with p-dimethylaminobenzaldehyde
(Ehrlich reagent) to produce colors ranging from light to
dark pink.
 Results are reported as Ehrlich units (EU), which are equal
to mg/dL,
 Chemstrip incorporates an azo-coupling (diazo) reaction
using 4-methoxybenzenediazonium-tetrafluoroborate to
o Vegetable peroxidase and bacterial enzymes, react with urobilinogen, producing colors ranging from white
including an Escherichia coli peroxidase, may also to pink. This reaction is more specific for urobilinogen than
cause false-positive reactions. the Ehrlich reaction.
 Results are reported in mg/dL.
BILIRUBIN
 Principle : Diazo Reaction
 Chemical Reaction:

 Appearance of bilirubin in the urine can provide an early

indication of liver disease
 Hepatitis and cirrhosis are common examples of conditions
that produce liver damage, resulting in bilirubinuria.

17
 o The Ehrlich reaction on Multistix is subject to a variety
of interferences, referred to as Ehrlich-reactive
compounds that produce false-positive reactions.
These include porphobilinogen, indican, p-
aminosalicylic acid, sulfonamides, methyldopa,
procaine, and chlorpromazine compounds. The
presence of porphobilinogen is clinically significant;
however, the reagent strip test is not considered a
reliable method to screen for its presence.
o The sensitivity of the Ehrlich reaction increases with
temperature, and testing should be performed at room
temperature.
NITRITE
 Principle: Greiss Reaction
 Chemical reaction :
 Provides a rapid screening test for the presence of urinary
tract infection (UTI)
 The chemical basis of the nitrite test is the ability of certain
bacteria to reduce nitrate, a normal constituent of urine, to
nitrite, which does not normally appear in the urine
 Nitrite is detected by the Greiss reaction, nitrite at an acidic
pH reacts with an aromatic amine (para-arsanilic acid or
sulfanilamide) to form a diazonium compound that then
reacts with tetrahydrobenzoquinolin compounds to produce
a pink-colored azodye.
 Results are reported only as negative or positive.
o Bacteria that lack the enzyme reductase do not
possess the ability to reduce nitrate to nitrite.
o Reductase is found in the gram-negative bacteria
(Enterobacteriaceae) that most frequently cause UTIs.
o Nitrite tests should be performed on first morning
specimens or specimens collected after urine has
remained in the bladder for at least 4 hours.
o Further reduction of nitrite to nitrogen may occur when
large numbers of bacteria are present, and this causes
a false-negative reaction.
o Large amounts of ascorbic acid compete with nitrite to
combine with the diazonium salt
LEUKOCYTE ESTERASE
 Principle : Hydrolysis of acid ester
 Chemical reaction:
 Increased urinary leukocytes are indicators of UTI.
 Neutrophils are the leukocytes most frequently associated
with bacterial infections
 Positive LE test result is most frequently accompanied by
the presence of bacteria, which, as discussed previously,
may or may not produce a positive nitrite reaction
 The reagent strip reaction uses the action of LE to catalyze
the hydrolysis of an acid ester embedded on the reagent
pad to produce an aromatic compound and acid. The
aromatic compound then combines with a diazonium salt
present on the pad to produce a purple azodye.
 Reactions are reported as trace, small, moderate, and large
or trace, 1+, 2+, and 3+.
 Lymphocytes, erythrocytes, bacteria and renal tissue cells
do not contain esterases.
o Highly pigmented urines and the presence of the
antibiotic nitrofurantoin may obscure the color reaction.
o Crenation of leukocytes preventing release of
esterases may occur in urines with a high specific
gravity.
SPECIFIC GRAVITY
 Principle : Change in pKa of polyelectrolyte
 Chemical Reaction:
 The polyelectrolyte ionizes, releasing hydrogen ions in
proportion to the number of ions in the solution. The higher
the concentration of urine, the more hydrogen ions are
released, thereby lowering the pH.
 Incorporation of the indicator bromthymol blue on the
reagent pad measures the change in pH. As the specific
gravity increases, the indicator changes from blue (1.000
[alkaline]), through shades of green, to yellow 1.030 [acid]).
Readings can be made in 0.005 intervals by careful
comparison with the color chart.
18
 o The reagent strip specific gravity measures only ionic
solutes.
o Specimens with a pH of 6.5 or higher have decreased
readings caused by interference with the bromthymol
blue indicator (the blue-green readings associated with
an alkaline pH correspond to a low specific gravity
reading).
CHEMICAL EXAMINATION: PART 2
CLINICAL SIGNIFICANCE AND CONFIRMATORY
TEST
PH
 Clinical Significance:
 Along with the lungs, the kidneys are the major regulators
of the acid–base content in the body.
o Secretion of hydrogen in the form of ammonium ions,
hydrogen phosphate, and weak organic acids, and by
the reabsorption of bicarbonate.
 Healthy individual
o First morning specimen with a slightly acidic pH of 5.0
to 6.0
o Normal random samples can range from 4.5 to 8.0.
o pH of freshly excreted urine does not reach above 8.5
in normal or abnormal conditions.
 Renal calculi formation
o Most common constituent of renal calculi is calcium
oxalate.
 Treatment of UTI
o valuable in treating urinary tract infections caused by
urea-splitting organisms
 Precipitation
o Medications: methenamine mandelate (Mandelamine)
and fosfomycin tromethamine (Monurol) are
metabolized to produce an acidic urine.
PROTEIN
 Clinical Significance:
 Protein test is the most indicative of renal disease.
 Normal urine contains very little protein less than 10 mg/dL
or 100 mg per 24 hours is excreted.
 Albumin is the major serum protein found in normal urine.
o Other proteins: serum and tubular microglobulins,
Tamm-Horsfall protein (uromodulin), and prostatic,
seminal, and vaginal secretions.
 Clinical proteinuria is indicated at 30 mg/dL or greater (300
mg/L).
 Prerenal
o Conditions affecting the plasma prior to its reaching the
kidney.
o Caused by increased levels of hemoglobin, myoglobin,
and the acute phase reactants.
o Multiple myeloma is a cancer of plasma cells. Plasma
cells make the antibodies (also
called immunoglobulins)
 Bence Jones protein by persons with multiple
myeloma.
o prerenal proteinuria is usually not discovered in a
routine urinalysis.
 Renal
o Glomerular or tubular damage.
o Preeclampsia is often precluded by gestational
hypertension.
o Diabetic nephropathy is common with both type 1 and
type 2 diabetes mellitus.
o Orthostatic proteinuria, or postural proteinuria.
Frequent in Young adults.
 Tubular
o Fanconi syndrome is a rare disorder of kidney tubule
function that results in excess amounts of glucose,
bicarbonate, phosphates (phosphorus salts), uric acid,
potassium, and certain amino acids being excreted in
the urine.
 Post renal
o Protein can be added to a urine specimen as it passes
through the structures of the lower urinary tract
OTHER TEST: SSA
 The sulfosalicylic acid (SSA) test is a cold precipitation test
that reacts equally with all forms of protein.
 All precipitation tests must be centrifuged to remove any
extraneous contamination.
GLUCOSE
 Clinical significance:
19
  Reducing sugars, including galactose, lactose, fructose,
maltose,pentoses, ascorbic acid, certain drug metabolites,
and antibiotics such as the cephalosporins.
 Clinitest does not provide a confirmatory test for glucose.
 Clinitest tablets are very hygroscopic and should be stored
in their tightly closed packages.
 A strong blue color in the unusedtablets suggests
deterioration due to moisture accumulation, as does
vigorous tablet fizzing.

KETONES
 Clinical significance:
 Glucose filtered by the glomerulus is actively reabsorbed in
the proximal convoluted tubule.
 Tubular reabsorption of glucose is by active transport.
 (renal threshold) for glucose is approximately 160 to 180
mg/dL
 Fasting prior to the collection of samples for screening tests
is recommended.
 Testing for urinary ketones is most valuable in the
 Hyperglycemia Associated management and monitoring of insulin-dependent (type 1)
o Hyperglycemia of nondiabetic origin. diabetes mellitus.
 Pancreatitis, acromegaly, Cushing syndrome,  Increased accumulation of ketones in the blood leads to
hyperthyroidism, pheochromocytoma, and electrolyte imbalance, dehydration, and, if not corrected,
thyrotoxicosis. acidosis and eventual diabetic coma.
 primary function of insulin (glycogenesis), these
opposing hormones (glycogenolysis) OTHER TEST: ACETEST
 Epinephrine is also a strong inhibitor of insulin
secretion and is increased in severe stress,
cerebrovascular trauma and myocardial infarction.

 Renal Associated
o Referred to as “renal glycosuria”
o End-stage renal disease, cystinosis, and Fanconi
syndrome.

OTHER TESTS: CLINITEST

 Acetest (Siemens Healthcare Diagnostics Inc., Deerfield,
IL) provides sodium nitroprusside, glycine, disodium
phosphate, and lactose (Color differentiation) in tablet form.
 If the specimen is not completely absorbed within 30
seconds, a new tablet should be used.

BLOOD
 Clinical Significance:

 The test relies on the ability of glucose and other BILIRUBIN

substances to reduce copper sulfate to cuprous oxide in the  Clinical Significance:
presence of alkali and heat.
 The tablets contain copper sulfate, sodium carbonate,
sodium citrate, and sodium hydroxide.

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 NITRITE
 Clinical Significance:

LEUKOCYTE ESTERASE
 Clinical significance:

SPECIFIC GRAVITY:
 Clinical Significance:

 Red blood cells is approximately 120 days

 Destroyed in the spleen and liver by the phagocytic cells of
the reticuloendothelial system.
 Liberated hemoglobin is broken down into its component
parts: iron,protein, and protoporphyrin.
 The bilirubin is then released into the circulation, where it
binds with albumin and is transported to the liver.
 In the liver, bilirubin is conjugated with glucuronic acid by
the action of glucuronyl transferase to form water-soluble
bilirubin diglucuronide.
 Passed directly from the liver into the bile duct and on to the
intestine. Intestinal bacteria reduce bilirubin to urobilinogen.

OTHER TEST: ICTOTEST

 P-nitrobenzene diazonium-p-toluenesulfonate
 SSA
 Sodium Carbonate
 Boric acid

UROBILINOGEN
 Clinical significance:

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