AUBF (MIDTERMS)
semipermeable membrane. A method of cleansing the
PREGNANCY TEST
CLASSIFIED INTO FOUR (4)
● Biological PT
○ Uses test animals
○ The most accurate in producing results.
 Immunological PT
 Chemical PT
 Clinician's Test
The main purpose of PT is to determine the normal
pregnancy.
➔ The woman becomes pregnant with the development of the
embryo coming from the union of egg and sperm. Later on, it
becomes a fetus and when delivered, it is called a baby.
➔ Period of 9 months
➔ PT is also done to differentiate true pregnancy from other
conditions such as hydatidiform mole (latu latu - agar).
➔ Even if it is not a fetus, the feeling of the female would be as
if they are pregnant
(cravings, nausea and vomiting).
➔ In the case of male patients, it is to determine whether they
have testicular embryonal carcinoma (teratoma). Also tumor
of the pituitary gland.
The principle depends upon the hormone, hCG. can be found in
blood in highest concentration. Use specimens with 2nd highest
concentration as blood extraction is refused. It is urine.
THE HORMONE CAN ALSO BE FOUND IN
● (CSF) Cerebrospinal fluid
● Saliva
● Sweat
● Tears
● Friedmann’s Test
Measurement of the hormone is in international units or mouse unit.
Gonadotropin activity of 0.1mg of hCG preparation required to
produce cornification of the vaginal epithelium of an immature
female rat.
 2 lines = positive
 1 line = negative
If positive with males, (false positive) patients may have abnormal
conditions, for females hydatidiform mole.
● Patient should avoid drugs with hCG
● or with toxic effects or tested with test animals.
If urine is alkaline, it might kill the test animal.
Death of test animal may be due to:
● Toxic Substances
● Alkaline Urine
● Bacterial Contamination
As a remedy to be able to repeat the test:
● Detoxification of urine.
● Addition of acid in the form of hydrochloric acid
with concentration of 0.1m. ph is 6.0.
● Dialysis - urine is placed in a cellophane bag
and is immersed with clean running water. Dialysis is
separation of solid from liquid portion using a
urine (ie crystalloids are removed).
RESULTS
Aside from the true pregnancy, we can obtain false positives.
ECTOPIC PREGNANCY
the fetus did not develop in the uterus, ie in the fallopian tube
('dating place' of sperm and egg). After fertilization in the fallopian
tube, the fetus is stuck.
● Physicians suggest abortion. if carried for 9 months.
● Sometimes the baby survives, sometimes both die.
CAUSE OF FALSE POSITIVES:
● When the female patient has increased
FSH (follicle stimulating hormone).
● When a female patient has appendicitis.
● Has incomplete abortion - there are certain traces of
tissues left in the uterus (endometrial lining). Can result in
bleeding. Do the DNC. Dilatation and curettage.
-seasonal variation of test animals
-if the test animal is pregnant, the natural result is positive.
BIOLOGICAL PREGNANCY TEST
Aschheim-zondek
● Use of white mice
● Immature female white mice. age is estimated. should be
21-30 days old.
● Quantitative test
● Purpose is to differentiate true pregnancy from
chorioepithelioma.
● A 24 hour urine sample is being injected into the
subcutaneous of the white mice. After about 48 hours,
sacrifice the mice and do an autopsy. Dissect the animal
and observe the ovary of the test animal. Observe corpora
hemorrhagica (red spots), Corpora lutea (yellow spots).
This indicates a positive result.
● Quantitative test
● Used for suspected hydatidiform, or teratoma.
● Principle is when hCG is injected to a female
virgin rabbit, it will produce 1-6 or more corpora
hemorrhagic on the surface of the ovary. When the ovary
becomes full, we can observe corpora lutea.
In the lab, female and male cages are separated. Animals are
born there and as they are separated, one will know if animals
are virgins or not. As for the rabbit, observe the behind. If shaped
like a v, it is a virgin. if shaped like a u, it is used.
Hoffmann’s Test
● Also makes use of female virgin rabbit
● Specimen is serum
● Injected into the marginal ear of the rabbit
● Same result observation in Friedmann's
Test.
Frog Test
Two (2) Types
1. Hogbem's Test
● using female frog
Principle:
when hCG is injected into a female frog, it will cause excretion of
eggs or ova. The urine is injected into the thigh muscle of the female
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frog. Set aside the frog for about 8-12 hours. During this period, do
not feed the frog. 12 hours after the urine of the frog is collected,
using a syringe without a needle, position in the urethra of the frog
and extract. Urine is placed on the glass slide. Look for eggs or ova
of the frog.
2. Gallimainini Test
● using male
Principle:
When hCG is injected to a male frog, it causes the release of cigar
shaped spermatozoa. Same procedure as above.
If there is no hCG in the patient sample, there is no egg or sperm
produced by frogs
 Xenopus laevis (South african frog) - female frog
 Rana vittigera and Rana pipiens (green grass
frog) - male frogs
CHEMICAL PREGNANCY TEST
➔ Less accurate, less popular
➔ Mixture of certain reagents to the urine sample.
Pregnanediol Test
 Detects progestene.
 Color is observed, deep orange to
dark brown is positive.
 Few amounts are added.
Iodine or Lugol’s Reaction Test
● Detects the presence of epinephrine.
● Positive is deep orange to deep brown.
Cytotrophoblast cell of the placenta produces hCG hormone. Found
in highest concentration in serum, 2nd in urine and next in CSF.
CHEMICAL PREGNANCY TEST
Cannot be done outside the physician’s clinic
Prostigmin Withdrawal Bleeding Test ??
● Prostigmin is injected to the patient, if a woman is going to
menstruate 2 weeks after the procedure, she is not pregnant.
Skin Colostrum Test
● First secretion of milk by the mammary glands.
● Colostrum is injected subcutaneously in the forearm of the
patient.
● Similar procedure to antibiotics. Small amount of it should
be injected to observe for allergic reaction.
● If the area swells and shows reddish coloration, not
pregnant.
● If there is no swelling and reddish coloration, she is
pregnant.
➔ Colostrum is secreted only by females who are pregnant or
delivered. If not, colostrum is a foreign substance to the body and
allergic reactions will manifest.
Primodos oral
● Comes in the forms of tablet
● Physician gives two tablets
● Taken once a day for two consecutive days
● Total of four tablets
● Observe five days after. If a woman menstruates, she is
not pregnant. If she does not menstruate, she is pregnant.
Gestex
● These tablets are taken in the same manner as primodos
oral.
● Only one tablet is taken for a day only.
● After 45 days, if the patient menstruates, she is not
pregnant.
Back in the olden days, there were many women who had wrong
interpretations about these procedures. They thought that gestex
would abort the baby.
IMMUNOLOGICAL PREGNANCY TEST
Makes use of antigen or antisera, same procedure in blood typing.
➔ Kit: vial of antisera, dropper (sometimes there is filter inside),
slide (sometimes made of glass or glossy material similar to
glossy camera film).
➔ Drop of antisera is dropped next to the drop of urine, then
mixed by swirling the slide.
➔ If agglutination occurs, positive for direct AT
➔ Indirect AT: if there is agglutination, negative. If not, positive.
➔ Specimens should be freshly voided. Not refrigerated, not
preserved.
➔ Antigen/antisera should be refrigerated. Before using vial
should be rubbed with palms
➔ IPT is not 100% accurate
➔ The IPT result will show in about 2 minutes.
➢ Biological is most popular as it is accurate.
In the Mid and late 90s, the test kit was hexagonal. There is a
specific area for drop of urine. There will be a plus sign observed
for positive and minus sign for negative.
A patient who would undergo a pregnancy test
should avoid drugs with hCG or other substances that may affect
the result.
(CSF) CEREBROSPINAL FLUID
 Not collected by medtech; only those who are trained
 Clear and colorless as distilled water
➔ NEEDLE used is not same as the needle in blood
collection (Needle is much longer)
➔ COLLECTION is done in between the 3rd or 4th Lumbar
Bone (between the SPACE, not on the Bones or Veins).
 Choroid Plexus – between the 3rd and 4th lumbar space.
pH of CSF
 Normally slightly Alkaline (7.30 – 7.45)
Specific gravity
 1.006 – 1.008
Specific gravity
 Normally Clear and Colorless, akin to distilled water.
 If there is a Bright Red Color (Fresh Blood) – it might due
to puncturing of a vessel (vein) during the insertion of the
needle.
 If there is a Brownish Red Color (Old blood) – there
might be intracranial hemorrhage.
 If the color is Yellow Xanthochromic – it might be due
blood pigments (RBC disintegration).
 If the color is Greenish/ Grayish – it is due to pus cells;
severe inflammatory reaction; acute meningitis.
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  After collecting the CSF in the syringe, it will be placed
Note:
into 3 labelled tubes.
 Transparency of CSF = Clear
Tube 1 (Base): Use for Chemical and Serological
 Number of WBC < 200 /mm3
Tests.
Tube 2 (Middle): Use for Microbiological
Coagulation examination (Free of microorganisms).
 CSF does not usually coagulate.
Tube 3: Use for Cell count – Wherein, immediate
 But whenever coagulation is observed, it is “clinical
processing is required since WBCs and RBCs begins
significant”.
to lyze within 1 hour. But, if the specimen cannot be
 If the coagulation is described as “Kabuet” (pine tree or
analysed, it has to be Refrigerated. Since, if the
web like); it indicates Tuberculosis meningitis.
specimen is left standing for about 2 hrs, about 40%
 If there is heavy coagulum + sediments occur; it is seen in
of WBCs will disintegrate.
Acute suppurative meningitis.
Xanthochromia
 Complete coagulation with Xanthochromia but without - Describing the color of the specimen as yellow, pink, or
hemorrhage; occurs in Froin’s syndrome (refers to spinal orange.
sub-arachnoid block).
 Test to confirm the presence of Sub-arachnoid block is Factors that cause Xanthochromia
known as Queckenstedt Test. 1. Present of RBC degradation products.
2. If the color is PINK, it has a very slight amount of
Oxyhemoglobin.
Prodcution of CSF 3. If the color is ORANGE, it has a heavy Hemolysis.
 CSF is produced in the Choroid Plexuses of the Lumbar 4. If the color is YELLOW, it indicates the conversion of
ventricles and the 3rd and 4th ventricles of Lumbar region. Oxyhemoglobin to Unconjugated bilirubin.
 Usually, 20 ml of CSF is produced by the Adult every 5. Elevated serum bilirubin, carotin, increase protein
hour. concentration, and melanoma pigment.
 Adult produces 20 – 150 ml of CSF.
 Infants produces 10 – 60 ml of CSF. NOTE
 The present of increase bilirubin signifies IMMATURE
NOTE LIVER FUNCTION.
➔ Backbone is joined together by Fibro (fibrous)  When the CSF is grossly bloody, Intracranial
cartilage. Whereas, the backbone has 5 lumbar bones. hemorrhage is observed.
➔ Skull has a layer of CSF that protects the Brains.  If there is really a hemorrhage, there is a way to determine
However, in collection of CSF, skull is the Last option. whether it is resulting from a hemorrhage or just from a
 CSF specimen should be treated and handled traumatic tap (insertion of the needle to the lumbar
carefully as; space), there are 3 Visual examinations.
1. It might contain organisms that can cause 1. Distribution of Blood =
disease (infectious). a. Uneven distribution = Traumatic Tap
2. It is not easily collected. b. Even distribution = Cerebral hemorrhage
➔ In the Brain there are 3 layers. 2. Clot Formation
I. Dura mater – Outer layer, lining the skull and the a. With Clot Formation = Traumatic Tap
vertebral canal. b. NO Clot Formation = Cerebral (intracranial)
II. Arachnoid mater – inner membrane. hemorrhage
III. Pia mater – thin membrane that lies the surfaces of 3. Xanthochromic Supernatant
the brain and spinal cord. - Hemolysis becomes noticeable when RBC remain in the
Collection of CSF CSF for at least 2 hrs.
 CSF specimen is always a priority (STAT), as it does not - For the recent hemorrhage, will produce a clear
contain enough Fibrinogen to form a clot. supernatant and when there is introduction of serum
 However, if the CSF is not in a STAT basis such as for the protein it may cause xanthochromic specimen.
following; Preservation of CSF
a. Hematologic exam - the tube has to be  Sodium Fluoride - prevent glycolysis by inhibiting the
Refrigerated. enzymatic activity (1 mg of fluoride per mL of CSF).
b. Microbiology – tube has to remain in a  1 mg NaF / 1 ml of CSF (1:1)
room temperature. Macroscopic Examination
c. Chemistry and Serology – tube has to - Measure the volume of the specimen (especially body
be frozen. fluids)

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 - Normal Volume of CSF = 1cc / lb (pound) of body  Reporting should be in order (sequential)
weight Interpretation:
- Daily production of CSF volume is roughly from 100 – 150 Unchanged Deep Red – 0
cc. Red to Blue – 1
Lilac to Purple – 2
NOTE Deep Blue – 3
 Increase amount of the CSF volume is observed in acute
Pale Blue – 4
and chronic congestion of the meninges (membrane).
Colorless – 5
 Increase amount in acute and chronic infections,
especially when there is production of Exudates (liquids NOTE
with inflammation) and Transudates (liquids without  Normal Reaction – all tubes (1-10) is unchanged.
inflammation process).  Paretic Curve (Zone 1) – 1st 3 tubes has Increase
number (ex. 5, 5, 5, 5, 5, 4, 3, 0, 0, 0). It is seen in
Chemical Examination for CSF insane and multiple myeloma patients.
➔ Qualitative Tests for Proteins (specifically for  Leutic / Tabetic Curve (Zone 2) – (ex. 0, 0, 0, 3, 3,
GLOBULINS): 3, 3, 0, 0, 0). It is seen in syphilitic patients.
1. Nonne - Apelt Test (Globulins)  Meningitis (Zone 3) – (ex. 0, 0, 0, 0, 2, 4, 4, 4, 4, 0).
2. Ross - Jones Test (Globulins) It is seen in Tuberculosis meningitic patients.
3. Pandy’s Test (Albumin and Globulins)  Curve in Brain tumor and Tuberculosis
4. Noguchi’s Test (Protein) meningitis patient – (ex. 0, 0, 0, 0, 2, 4, 3, 3, 3, 3).
5. Benedict’s Test (Glucose/Sugar)
Quantitative Tests: NOTE

1. Nelson – Somogyi Test (Glucose / Sugar)

- Use in Clinical Chemistry  Paretic curve shows the marked change in the 1st 3

2. Schales and Schales Test tubes.

- Test for Chlorides  Leutic curve shows the marked change in 3rd and 4th
3. Lange’s Colloidal Gold Test tube.
- Albumin / Globulin Ratio Test  Meningitis curve shows the marked change in the last 3
- Uses 10 test tubes to make dilutions. tubes.
Procedure/s:
o 10 test tubes are needed + 2 tubes (as control).
o on the 1st tube, 0.9 cc of 0.4% sodium chloride
solution is placed.
o Then, 0.5 cc of the solution is transferred into
the other tubes up to the 11th tube.
o Place 0.85 cc of 1% NaCl in the 12th tube.
o Add 0.1 cc of CSF in the 1 st tube then mixed it
well by sucking the fluid into the pipette (4x).
o After mixing, 0.5 cc will be transferred to the 2 nd
tube (repeat in each successive tube including
the 10th tube).
PREGNANCY TEST
o But the 11th and 12th tube are not included.
o to each of the 12 tubes, add 2.5 cc of colloidal
gold solution. .

NOTE
 11th tube serves as a control for the Stability of the
colloidal gold.
 12th tube serves as a control for the Sensitivity of the
colloidal gold. It should become colorless within an hour.

 Let it stand overnight and read the result the next morning.
In which, color changes should be observed.

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