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Aubf - Prelims
Aubf - Prelims
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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.
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.
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Specific requirements of this OSHA standard include the CHEMICAL HANDLING
following: Chemicals should never be mixed together unless specific
o 1. Requiring all employees to practice UP/Standard instructions are followed and they must be added in the
Precautions order specified.
o 2. Providing PPE to employees
o 3. Providing sharps disposal containers and prohibiting CHEMICAL HYGIENE PLAN
recapping of needles Required by OSHA
o 4. Prohibiting eating, drinking, smoking, and applying
cosmetics, lip balm, and contact lens in the work area Purpose of the plan:
o 5. Labeling all biohazardous material and containers o 1. Appropriate work practices
o 6. free immunization for HBV o 2. Standard operating procedures
o 7. daily disinfection protocol for work surfaces o 3. PPE
o 8. Providing medical follow-up for employees who have o 4. Engineering controls (fume hoods, safety cabinets,
been accidentally exposed to blood-borne pathogens etc…)
o 9. Documenting regular training in safety standards for o 5. Employee training requirements
employees. o 6. Medical consultation guidelines
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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.
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.
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
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.
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
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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.
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RENAL BLOOD FLOW Podocytes are the intertwining foot processes of the inner
The renal artery supplies blood to the kidney. layer of Bowman’s capsule.
Approximately 25% of the blood from the heart goes to the The barrier contains a shield of negativity that repels
kidney. molecules with a positive charge.
Blood enters the capillaries of the nephron through the CELLULAR STRUCTURE OF THE GLOMERULUS
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
CAPILLARY TUFT
collectively term for eight capillary lobes
BOWMANS CAPSULE
Beginning of the renal tubule.
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Active transport, like passive transport, can be influenced
by the concentration of the substance being transported.
(Tm) - maximal reabsorptive capacity
Must to know about glomerular filtration: The plasma concentration at which active transport stops
o 1. Every minute approximately two to three million is termed the renal threshold.
glomeruli filter passes the glomerulus. Knowledge of the renal threshold and the plasma
o 2. This filtration is nonselective and can be concentration can be used to distinguish between excess
differentiated to plasma by the absence of the plasma solute filtration and renal tubular damage.
proteins
o 3. Analysis of the fluid as it leaves the glomerulus TUBULAR CONCENTRATION
shows the filtrate to have a specific gravity of 1.010 and
DLH and ALH IS THE BEGINNING OF RENAL
confirms as an ultrafiltrate of plasma.
CONCENTRATION because the filtrate is exposed to the
high osmotic gradient of the renal medulla.
TUBULAR REABSORPTION Water is removed by osmosis in the descending loop of
The body cannot lose 120 mL of water-containing essential Henle.
substances every minute. Sodium and chloride are reabsorbed in the ascending loop.
Proximal convoluted tubule is the one responsible of
reabsorbing most of the essential substances and water OSMOSIS
needed by the body.
A process by which molecules of solvent tend to pass
through a semipermeable membrane from a less
concentrated solution into a more concentrated one.
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
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Normal plasma creatinine range: 0.5-1.5mg/dl 2. PHENOLSULFONPHTHALEIN
NOT CURRENTLY PERFORMED
GLOMERULAR FILTRATION TESTS STANDARDIZATION AND INTERPRETATION OF
CREATININE CLEARANCE DISADVANTAGES RESULT IS DIFFICULT
Can be secreted by the tubules COLORLESS IN ACID SOLUTION AND RED IN
Secretion is affected by medication ALKALINE SOLUTION
Bacteria will break down urinary creatinine
Dietary 3. URINARY AMMONIA AND TITRATABLE ACIDITY
Cannot be used in patient with muscle-wasting disease / DETERMINES THE DEFECTIVE FUNCTION IN THE
athletes ABILITY OF THE KIDNEY TO PRODUCE ACIDIC URINE.
It needs correction for body surface area especially in FACTORS INVOLVE TO PRODUCE AN ACIDIC URINE
children.
o Newer methods that do not require the collection of A. TUBULAR SECRETION OF HYDROGEN ION
timed (24-hour) using serum creatinine, cystatin c or B. DCT PRODUCTION AND SECRETION OF AMMONIUM
beta2-macroglobulin values. IONS
A NORMAL PERSON EXCRETES 70meq/day of acid in
MEDICATIONS the form of either titratable acid, hydrogen phosphate ions ,
Gentamicin ammonium ions.
Cephalosporin A diurnal variation affects urine acidity
Cimetidine ( Tagamet) inhibit tubular secretion of creatinine Renal tubular acidosis is the inability to produce an acidic
estimated glomerular filtration rate (eGFR) 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
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.
OSMOMETRY
Test use for quantitative measurement of renal
concentration ability.
Reported in milliosmole (mOsm)
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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
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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
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
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ODOR
It is not part of routine urinalysis.
Odor Cause
Aromatic normal
Mousy Phenylketonuria
Rancid Tyrosinemia
Bleach Contamination
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