Gastric Fluid

Significance
• Determines whether or not a patient can secrete gastric fluid
• Measures amount of gastric acid that can be secreted by one with
ulcer symptoms
• Help determine the disturbed function of the GI system
Cells of the Stomach
1. Parietal cells – produces HCl and Intrinsic factor
*HCl – converts pepsinogen to pepsin that catalyzes the
digestion of protein
*Intrinsic factor – responsible for Vitamin B12 absorption
2. Chief cells – produces pepsinogen that will be converted to pepsin
whenever HCl is present
3. Specialized G cells - produces Gastrin that stimulates parietal cell to
produce HCl.
Specialized G cells  Gastrin
Gastrin stimulates Parietal cells
HCl

Parietal cells HCl pepsinogen ------ > pepsin = digestion

 Intrinsic Factor  Vit. B12 absorption
Specimen collection
Gastric juice is obtained by insertion of a gastric tube into the stomach
Gastric tubes:
A. Levin tube = passed thru the nose
B. Rehfuss = passed thru the mouth
C. Disposable plastic tubes – usually employed

Specimen = fasting specimen, few ml to 50ml. Average of 30ml

Normal appearance of gastric specimen: Pale gray with mocus and no
food particles
Types of specimen
Basic acid output (BAO)
1 hr collection (4 fifteen minutes specimens)
Requires 12 hour fasting
No gastric stimulant needed

Maximum acid output (MAO)

1 hr collection (4 fifteen minutes specimens)
With gastric stimulants
Gastric stimulants
1. Test meals
a. Ewald’s meal: Bread and water/weak tea
b. Boa’s meal: Oatmeal (for lactic acid detection)
c. Riegel’s meal: Mashed potatoes, broiled beefsteak, bouillon
2. Chemicals
d. Pentagastrin
e. Histamine
f. Histalog
g. Insulin – assess successful vagotomy procedure
3. Sham Feeding
h. Fictitious feeding
i. Sandwich is chewed and then spit out.
Diagnex Blue Test / Tubeless Test
Specimen: Urine
Principle: an ion exchange resin (Amberlite cation) resin, coupled with a
dye, azure blue, is given by mouth after caffeine stimulation. In the
presence of free HCl, the azure blue is released from combination with
the resin in exchange for hydrogen ions. The azure blue is rapidly
adsorbed from the intestines and travels in the blood to the kidneys
and is excreted in urine. The appearance of azure blue is then an
indication that free HCl is present in the stomach.
Stimulants used: Test meals (Henry’s), Histamine(other books)
A. Zollinger Ellison Syndrome
Elevated gastrin levels
Elevated BAO/MAO results(highest elevation)
B. Pernicious Anemia
Shows zero BAO/MAO results
Achloridia (absence of free HCl)
Euchlorydria Normal free HCl
Hyperchlorydria Increased free HCl Peptic ulcer
Hypochlorydria Gastric fluid pH >3.5 but Carcinoma of the stomach
falls after gastric
stimulation
Decreased free HCl
Achloridria Gastric fluid pH >3.5 and Pernicious anemia
does not fall after gastric
stimulation
Absence of free HCl
Qualitative Test for Free HCl
Dimethylaminoazobenzol Reagent: alcohol solution = (+) cherry red
Gunzberg’s Reagent: phloroglucin, Vanillin, Alcohol = (+) purple red color
Boas Reagent: resorcinol, cane sugar, alcohol = (+) purple red color
Quantitative Test for Gastric Acidity
Free HCl Topfer’s method
 Titrate with NaOH
 pH indicator: dimethyl aminoazobenzol
 Endpoint: Canary Yellow
 Normal range: 25-50 degrees or 0.1 or 0.2 HCl
Total acidity  Titrate with NaOH
 pH indicator: phenolphthalein
 Endpoint: Faint pink
 Normal range: 50-75 degrees
Combined HCl(bound to CHON)  Titrate with NaOH
 pH indicator: sodium alizarin
 Endpoint: Violet
 Normal range: 10-15 degrees
Lactic Acid Test
Test Reagents Endpoint
Modified Uffelman’s FeCl3 + Phenol Yellow
Strauss FeCl3 + ether Yellow
Kelling’s FeCl3 Yellow
Vaginal Secretion
Vaginal secretions are examined in the clinical laboratory to diagnose
infections and complications of pregnancy, and for forensic testing in
sexual assault patients.

Vaginitis is one of the most common conditions diagnosed by health-care

providers for female patients, particularly of childbearing age. It is
characterized by abnormal vaginal discharge or odor, pruritus, vaginal
irritation, dysuria and dyspareunia.
Although symptoms for the various syndromes of vaginitis are similar, the
effective treatment for each depends on an accurate diagnosis. To
determine which is which, a careful microscopic examination is necessary.
Microscopic methods include saline wet mount examination, KOH
examination and Gram stain, which is considered the gold standard.
Specimen Collection and Handling
The specimen is collected by swabbing the vaginal walls and vaginal
pool to collect epithelial cells along with the vaginal secretions using
one or more sterile, polyester-tipped swabs on a plastic shaft or swabs
specifically designed by the manufacturer.

Cotton swabs should not be used because cotton is toxic to Neisseria

gonorrheae.
Wood in a wooden shaft may be toxic to Chlamydia trachomatis and
calcium alginate can inactivate herpes simplex virus for viral cultures.
Specimens should be analyzed immediately but, if a delay in transport
or analysis is necessary, specimen handling is based on the suspected
pathogen.
T. vaginalis & N. gonorrheae – room temp to preserve motility
*Specimens for T. vaginalis should be examined within 2 hours of
collection
C. Trachomatis and HSV – refrigerated to prevent overgrowth of normal
flora
Color and Appearance
Normal vaginal fluid appears white with a flocculent discharge
Microscopically, normal vaginal flora includes a predominance of large,
rod-shaped, gram-positive lactobacilli and squamous epithelial cells.
Abnormal vaginal secretions:
Bacterial vaginosis – thin, homogenous white to gray discharge
Candida infections – white “cottage cheese” – like discharge
T. vaginalis – yellow green, frothy, adherent discharge
C. Trachomatis – yellow, opaque cervical discharge
Diagnostic Tests
pH testing
This test should be performed before placing the swab into saline or
KOH solutions
This helps to differentiate the causes of vaginitis.
pH 3.8 – 4.5 – normal / healthy
pH 4.5 – vulvovaginal candidiasis
pH >4.5 – bacterial vaginosis, trichomoniasis, desquamative
inflammatory vaginitis and atrophic vaginitis.
Lactic acid provides an acidic vaginal environment with a pH value
between 3.8 – 4.5. This acidity suppresses the overgrowth of infectious
organisms such as Mobiluncus, Prevotella and Gardnerella vaginalis.
Some lactobacilli subgroups also produce hydrogen peroxide, which is
toxic to pathogens, and helps keep the vaginal pH acidic to provide
protection from urogenital infections.
Estrogen production also is necessary to preserve an acidic vaginal
environment.
Microscopic Procedures
Vaginal infections are usually diagnosed from microscopic examination.
Saline wet mounts and KOH mounts are the initial screening tests while
Gram stain is used as a confirmatory test for yeast/bacterial vaginosis.
2 glass slides are usually ordered.
3 glass slides if a Gram stain is requested.
Labels:
Name of the patient
Unique identifier e.g. control number
Squamous epithelial cells
They contain a prominent centrally located nucleus.
Large amount of irregular cytoplasm
Lacking granularity
Distinct cell margins/ borders
Clue cells
Large amount of irregular cytoplasm
Granular
INdistinct cell margins/ borders
WBC
Normal: rare to scanty numbers in vaginal secretion. Up to 2+

Abnormal: 3+ WBCs in vaginal secretion.

Significance: vaginal candidiasis, atrophic vaginitis or infection with T.
vaginalis, N. gonorrheae, Herpes simplex virus.
RBC
Confused with yeast cells and are distinguished from yeast cells by KOH,
which will lyse RBCs but allow the yeast cells to remain intact.
Parabasal Cells
The nucleus to cytoplasm ratio is 1:1 to 1:2, with marked basophilic
granulation or amorphic basophilic structures (blue blobs) in the
surrounding cytoplasm.
Increased number of parabasal cells along with large numbers of WBCs,
can indicate desquamative inflammatory vaginitis(DIV)
Basal cells
Basal cells are distinguished from WBCs that are similar in size by their
round rather than lobed nucleus.

Increased number of basal cells along with large numbers of WBCs, can
indicate desquamative inflammatory vaginitis(DIV)
Trichomonas Vaginalis
Has four anterior flagella and an undulating membrane that extends
half of the length of the body. An axostyle bisects the trophozoite
longitudinally and protrudes from the posterior end.
The “jerky” motion of the flagella and undulating membrane is a
characteristic of T. vaginalis can be observed in wet mount.
KOH preparation and Amine test
The KOH slide is prepared and the amine (Whiff) test is performed by
placing a drop of the saline specimen prepared from the collection
swab onto a properly labeled clean slide and adding 1 drop of 10% KOH
solution. The slide is then immediately checked for “fishy” amine odor.
The result is reported as positive or negative.
Other diagnostic tests
Gram stain – is considered the gold standard in identifying the
causative organism for bacterial vaginosis.
Scored gram stain = Nugent score

Nugent scoring:
0-3 = normal vaginal flora
4-6 = intermediate
>7 = bacterial vaginosis