Transudates and Exudates & Pregnancy Test PDF

LECTURE | MIDTERM Transcribed by: Faulien & Rhed | Organized by: Angela I. TRANSUDATES AND EXUDATES EXAMINATION MACROSCOPIC OR PHYSICAL EXAMINATION Point of Transudate Ex...

LECTURE | MIDTERM Transcribed by: Faulien & Rhed | Organized by: Angela I. TRANSUDATES AND EXUDATES EXAMINATION MACROSCOPIC OR PHYSICAL EXAMINATION Point of Transudate Exudate Transudates Difference 1. Color and 1. Clear and 1. Cloudy or Transudates are accumulated pathologic fluids of Appearance usually light purulent with non-inflammatory nature in the serous cavities of yellow, blood color the body. Usually, these are due to: colorless unless varies from 1. Cardiac failure in case of gray to light 2. Venous obstruction (portal vein) jaundice, when yellow 3. Hypoproteinemia (nephritis or necrosis) blood is depending on 4. Disturbance of circulation with passive present, it is the type of congestion and edema. amber or organism and yellow ir color density of the Transudates may develop in the: Pseudochylous fluid, example: 1. pericardium - hydropericardium fluid in chylous Green or 2. pleural cavity - hydrothorax but milky greenish yellow 3. abdominal cavity - ascitis appearance when infecting 4. scrotum - hydrocele due to lecithin organism is 5. joints – hydrarthrosis cholesterol. pseudomonas 6. sub-arachnoid space - cerebral edema. aeroginosa. Appearance may be serous EDEMA: or sero- When similar fluid develop from apparently the fibrinous as well same causes occurs in the tissues, due to as purulent. disturbance in the circulation. Mucopurulent and chylous Example: effusions. produced due to bacterial EXUDATES: origin. Exudates are purulent fluids that forms in any body cavity as a result of an inflammatory process. 2. Odor 2. Oderless 2. Odorless or The most common cause is bacterial invasion. It have a may be caused by parasite by a sterile irritating sickeningly agent such as bile or pancreatic juice. Exudate may sweet odor. be cloudy and serous, fibrinous, purulent, When infected hemorrhagic, chylous or chyloid or a combination by E. has a of these. fecal odor. Types of Exudates: 3. 3. Does not 3. Clots readily Serous — generally straw colored and contains very Coagulation coagulate because of the few cells. presence of fil rinogen Fibrinous - yellow in color and coagulates readily (Spontaneoue (spontaneously) due to rich fibrinogen content. coagulation) Purulent — contains abundance of pus cells, seen in pyogenic infection, pneumococeus and 4. Specific 4. Less than 4. More than staphylococcus infections. gravity 1.018 because 1.018 due to of low protein large amount Hemorrhagic - contains variable amount of blood. content. Not of protein Chylous - generally results from trauma and rupture more than 2.5 (albumin)), of a lymph vessel, due to destruction of the thoracic gm. % more duct. Milky in color and contains fat globules. Seen than 3 gm. % in filariasis and other environment of the lymph and WBC. vessels. Chyloid - contains lecithin with some fat and 5. Reaction 5. Alkaline 5. Acidic pseudoglobulin. Milky in color. Putrid - generally seen in gangrene. CHEMICAL EXAMINATION Serofibrinous - straw to yellow color containing I. ALBUMIN: proteins and seen in pleural effusion. A. Qualitative Serosanguinopurulent - bloody with pus cells. Use procedures similar to Urine Albumin Detection. yellowish to green and seen in hemorrhages, Color B. Quantitative is with streptococcal, staphylococcal infections. 1|Page Source: Book/notes of Ma’am Tan  LECTURE | MIDTERM Transcribed by: Faulien & Rhed | Organized by: Angela 1. Esbach's Method: 3. Ziehl-Neelsen Method Procedure: Difference Transudate Exudate Fill an Esbach's tube with the specimen up to the U Point of mark. 1. Crystals 1. No crystals, 1. May contain Add Esbach's reagent up to the R mark. if present, it crystals. Cholesterol Invert to mix. indicates associated with stasis. fatty degeneration. Add 0.1 to 0.2 gms. of barium sulfate (BaS04) or Hematoidin crystals Charcoal. (This is for absorption to increase rate of associated with sedimentation of albumin). hemorrhage. Stand the mixture for 15 to 20 minutes. 2. Cells 2. Few 2. Many cells are Read the amount of albumin present. mesothelial present with the II. SEROSAMUCIN: calls as well as predominance of: A. Qualitative blood cells a) PMN - suggests pyogenic 1. Rivalta's Test infection Procedure: b) Leukocytes suggest an Place a drop of acetic acid to 100 ml. water in a uncomplicated graduated cylinder. TB infection. a Allow a drop of fluid to fall in the solution. chronic Interpretation: pyogenic infection or Normal body fluids give no cloud certain blood Transudate gives a slight cloud - Negative disease. Exudate gives a heavy precipitate - Positive. c) Mesothelial - Suggest 2. Biuret Test transudate or May be used for protein determination in malignant transudates and exudates. The procedure is similar infection or with that of serum technique. disease. III. SUGAR CONTENT: d) Eosinophil- frequent Both the Qualitative and quantitative examinations tappings and are simi. lar to Urine. hypersensitivity. Eosinophils may Transudate - the sugar content is approximately occur in spinal same as blood sugar. fluids with or without Exudate - low sugar content as compared with eosinophi-lis, blood. reported in neu-rosyphilis or cysti carcosis of MICROSCOPIC EXAMINATION CNS. CYTOLOGIC EXAMINATION: The specimen should be fresh and citrated to prevent clotting. 3. Origin 3. Non- 3. Inflammatory A. Total Cells Count inflammatory Follow the same procedure as in CSF Total Celi 4. Chloride 4. Same as 4. Higher than Count. If the count is high, follow the WBC blood blood. procedure and make the necessary corrections. B. Differential Count Follow the same procedure as in CSF. Smear must SYNOVIAL FLUID be thin that the cells will not be disturbed and shrunken in appearance. I. SYNOVIAL FLUID Synovial fluid is obtained by aspiration of a joint, C. Bacteria bursa, or tendon sheath. Any of these stains may be used: II. CELL COUNT: 1. Wright's stain A. Method 2. Gram's Stain 2|Page Source: Book/notes of Ma’am Tan  LECTURE | MIDTERM Transcribed by: Faulien & Rhed | Organized by: Angela 1. Use the same method as described under spinal at the National Institute of Medical Research, fluid except, use 0.85% NaCL solution instead of London (First Standard). diluting fluid. International unit of chorionic gonadotropin was defined as the activity contained in 0,001279 mg 2. Normally there are 50-60 leukocytes per cu mm. of the Second International Standard. B. Differential Count International unit is the gonadotropin activity of 1. Centrifuge some of the fluid at high speed for 30 0.1 mg. of chorionic gonadotropin preparation minutes; pour off the supernatant fluid and make a which is required to produce cornification of the smear of the sediment. vaginal epithelium of an immature female rat. Mouse unit has the same definition but 2. When dry, make a Wright's stain and differentiate expressed in ce, of smallest dose or amount of 100 cells. urine expressed in ec. that will produce positive 3. Normal differential count reveals 60-70% result on, pregnancy. monocytes; 20-30% lymphocytes; 5-10% neutrophils; Chorionic gonadotropin is increased to a peak and 4% synovial cells. during the first two months after the first missed period (between eighth and the twelfth week of III. BACTERIOLOGY: amenorrhea, slowly decreases to a constant low A. Direct Smear level after the sixteenth week, and disappears 1. Centrifuge as described under differential count. during the first week after parturition. One-hour postpartum positive test but negative 3-4 days 2. Make a smear of the sediment, dry, fix, and make after delivery. a gram stain. In normal conditions of pregnancy wherein the 3. Examine for bacteria. urine has a very low titer (few amounts of hormone), a negative pregnancy test is B. Cultures detained. To obtain positive test, the small C. Complement-Fixation Test: amount of hormone may be concentrated by 1. To rule out syphilis, use synovial fluid in place of adding any of these substances: kaolin, absolute serum in the routine method for the Wassermann alcohol, and acetone (1 part urine to to 2 parts Test. acetones.) 2. To rule out gonorrhea, use synovial fluid instead of serum in the gonococcus Complement-Fixation IMPORTANCE OF PREGNANCY TESTS: Test. 1. To determine if a woman is pregnant or not, because some women need medications. II. PREGNANCY TEST 2. To determine if there has been complete or incomplete abortion. Pregnancy is a term used for a laboratory 3. To differentiate other pelvic diseases. examination for the confirmation of early normal pregnancy, evaluation of abnormal 4. To differentiate pregnancy from other pregnancy, and the study of trophoblastic uncommon condition like hydatidiform mole and diseases. All pregnancy tests were designed chorioepithelioma. to measure or detect human chorionic 5. To differentiate pathologic condition of male, like gona- dotropin (HCG), a hormone produced chorio-epithelioma, teratoma, choriocarcinoma of by viable placental tissue, the testis and semenoma. HUMAN CHORIONIC GONADOTROPIN: Human chorionic gonadotropin (HCG) is a PREPARATIONS OF A PATIENT FOR PREGNANCY TEST: hormone which is a glycoprotein with a 1. Fluid intake should be restricted from 3:00 p.m. molecular weight of approximately 50,000; it has until the morning urine has been collected. a relatively high carbohydrates content to the 2. Salicylates (aspirin), barbiturates, or other form of galactose (as much as 30% in some sedative should be avoided for at least 45 hours preparation), secreted by the cytotrophoblast prior to urine collection. cells of the placenta and are found in the blood, urine, amniotic fluid, colostrum, milk, fatal tissue 3. The first morning urine must be collected into a and nearly in all body fluids of a pregnant clean glass container. woman. Its level rises rapidly following 4. Collect about 60 ml. of specimen in order to have conception and becomes detectable as early sufficient amount for detoxification if in case as 10-14 days after conception (2 days after the necessary. missed period or 24th day after the last menstrual period (LMP). PREPARATION OF SPECIMEN: It is quantitated in international units (IU) of gonadotropin actively. 1. Specimen should be eleah; otherwise, it must be One international unit is defined as the specific filtered or centrifuged. gonadotropic activity of 0.1 mg. dried standard 2. No preservative should be added. 3|Page Source: Book/notes of Ma’am Tan  LECTURE | MIDTERM Transcribed by: Faulien & Rhed | Organized by: Angela 3. Specimen should be kept in the refrigerator and ovaries of rats, and secretion of sperm or ova in placed at room temperature for 30 minutes prior to toads and frogs. injection. The biologic effect of the hormone and the sensitivity of different animal species vary considerably. In amphibians there are also CAUSES OF FALSE POSITIVE REACTIONS: seasonal variations in sensitivity. 1. High titers of pituitary gonadotropin (PG) from any cause. 2. High titers of follicle-stimulating hormone (FSH) in meno- pause or primary ovarian failure. 1. Ascheim-Zondek Test 3. Variety of drugs such as chlorporomazine Materials: derivatives Com- pazine), phenothiazine a) 24-hour urine compound (Thorazine and Sparine), and aspirin should not be taken for at least 48 hours prior to test. b) 5 immature female white mice (7-10 gms. in weight and 18-23 days old) Alkaline must be 4. Seasonal variation of experimental animals (Frog acidified with glacial acetic acid (few drops). This is - positive in July and August). done because the hormone is active if the urine is 5. Ectopic pregnancy. acidic. 6. Acute appendicitis. Urine not tested right away are preserved in a 7. Incomplete abortions as long as placental tissue refrigerator. is present. Procedure: a) Inject 0.4 ml. of urine to each of the 5 mice CAUSES OF FALSE NEGATIVE REACTIONS: subsutaneously, three times daily for two days. Total 1. Low sensitivity of the animal, sometimes resulting dose is 2.4 ml. from seasonal variations or from metabolic b) Kill the mice 100-120 hours after the first injection depression by toxic urine. using ether. 2. Low titer or concentration of hormone. c) Autopsy the mice preventing the escape of 3. Toxic urine specimen. blood to the abdominal cavity since it will affect the coloring of the real effect. CAUSES OF TOXIC URINE: d) Positive result is shown by the enlargement of the 1. Excess amount of electrolytes, particularly every 2-3 times that of the normal size of ovary is pin potassium. head). 2. Bacterial contamination or other toxic Other distinguishing positive results are the yellowish substances. spots, the corpura lutea while the red spots are the 3. Salicylates and barbiturate medications. corpura hemorrhagica to graffian follicle from the corpura luteum. Still other evidences are the swelling or hyperemia of uterus and premature METHODS OF DETOXIFYING URINE: opening of vaginal canal. Test is positive in 95-98% 1. Dialysis in cellophane tubing for 30 to 60 minutes of early pregnancy cases. under running water may reduce the electrolyte Sensitivity: 1-6 IU/ml. content. 2. Acidification with 0.1 N hydrochorie acid to a pH of about 6.0 may make bacterially contaminated 2. Friedmann's Test urine less toxic. Materials: 3. Extraction with ether or alcohol. a) 15-20 cc. of urine. 4. Absorption with kaolin. b) Female rabbit (virgin rabbit) c) Weight over 4 pounds and more than 16 weeks MAIN GROUPS OF PREGNANCY TESTS: old. A. BIOLOGICAL PREGNANCY TESTS Procedure: These tests use animals and are considered the best for routine examination of pregnancy. They a) Inject 15 cc. of urine into the vein of the marginal are reliable and easy to perform and more ear of the rabbit. economical. b) Kill the rabbit after 30-48 hours after injection. c c) Positive result: Principle: Injection of human chorionic gonadotropin Ovaries are full, 1-6 or more corpura hemorrhagica (red spots projections at the surface of the ovary). (HCG) into certain animals produces characteristic anatomic changes, mainly as Yellowish spots or corpura lutea. Rossy spots are easily demonstrable luteinizing effect doubtful results (production of corpora lutea) in the ovaries of NOTE: mice and rabbits, pronounced hyperemia in the 4|Page Source: Book/notes of Ma’am Tan  LECTURE | MIDTERM Transcribed by: Faulien & Rhed | Organized by: Angela Positive results occur in 95-97% of normal pregnancy b. Kelso Test after 10-14 days of conception until the first Materials: trimester. a) Urine about 5 ml. False positive results in: b) Female virgin rat 30-65 gms and 3-6 weeks old. a) When rabbit is the old and is really pregnant itself. Procedure: b) In ectopic pregnancy, but negative in 3 weeks after onset of vaginal bleeding, after fetal death or a) Inject 4 ml. of urine subcutaneously into the rat. incomplete abor- tions, but positive as long as b) After a period of 24 hours, the animal is killed. c) placental tissue remains in uterus. An abdominal incision is made and the ovaries are c) In man in cases of: examined. 1) Choriocarcinoma or testicular embryonal c) If ovaries are small and pale, the test is negative. carcinoma, or teratoma. If the ovaries are enlarged and bloody, the test is positive. 2) Tumor of pituitary glands. Sensitivity: 1 IU/ml 3) Increase gonadotropin hormone. False negative result in: Kupperman Test When rabbit is too young, not able to produce a positive reaction. the ovary is narrow, flat, opaque Materials: in appearance, and uterine color is pure white. a) 5 ml urine Sensitivity: 10-15 IU/ml. b) Female virgin rat 22-40 days old and 30-65 gms. Procedure: 3. Hoffmann's Test a) Inject 4 ml. of urine intraperitoneally to the rat. (Similar to Friedmann's Test) b) After a period of 2-24 hours, the rat is killed. Materials: c) Examine for ovarian hyperemia for positive results. a) Female virgin rabbit, 4 pounds; 16 weeks old Sensitivity: I IU/ml. b) Serum d. The test is reliable and rapid. Procedure: a) Extract 12 cc. of whole blood from pregnant Frog Test: woman an allow the blood to clot or centrifuge to a. Female Frog Test or Hoghen's Test separate the serum from the cells. Principle: The mature female frog carries eggs b) Inject 3-4 ce. of serum in the vein of the marginal throughout the year hut deposit them only after ear of the rabbit. mating or after injection of pregnancy hormones. c) Kill the rabbit 30-40 hours after the injection. Materials: d) Positive result is similar to that of the Friedmann's a) Female frog (xenopus laevis and South African Test. clawed frog). b) Syringe and hypodermic needle, gauge 22. 4. Rat Ovarian Hyperemia Tests c) Alcohol and water. a) Frank-Berman Test d) Tissue paper. Materials: e) About 20 cc. of first morning urine. a) 20 cc. centrifuge urine b Procedure: b)2 immature female rats (50 gms., 3-6 weeks old) a) Inject 2 cc. of urine into the muscle of the thigh, Procedure: then towards the dorsal lymph sac subcutaneously. a) Inject subcutaneously 5 ce urine into each of 2 b) Set the frog aside for eight to twelve hours. Don't immature female rats. Space injection at least 4 feed the frogs during the test. hours apart. c) Positive result is shown by extrusion of eggs 8-12 b) Sacrifice animal by illuminating gas 16-24 hours hours after injection. after first injection. NOTE: Female frogs are extremely sensitive to many c) Dissect out ovaries and examine grossly with the drugs, example: morphine derivatives and naked eye. A negative ovary or slightly pinkish. barbiturates, so that even a small amount may kill Minute colorless folicles can be seen. A positive the frog. ovary is usually, but not always, enlarge and is entirely reddened (or may have many. distinct red Sensitivity: 75-100 IU/ml. spots) b. Male Frog Test or Galli Mainini Test Sensitivity: 1 IU/ml. 5|Page Source: Book/notes of Ma’am Tan  LECTURE | MIDTERM Transcribed by: Faulien & Rhed | Organized by: Angela Principle: After injection of urine extract or serum than in the winter. For this reason, monthly sensitivity containing HCG into the lymph sae of the male frog, tests should be performed. expulsion of spermator into the cloaca within 2 to 4 hours. B. IMMUNOLOGIC PREGNANCY TESTS: Materials: Due to the many disadvantages of the biologic a) Male frog (Rana vittegera, Rana pepiens green tests such as seasonal variation in sensitivity, grass frog. Rana clamitans, Bufo americanus, Bufo maintenance of animals and the time and bufo. expenses involved, the search for development of in vitro methods has been pursued. b) Syringe and hypodermic needle gauge 22. In 1960, four groups of researchers (independent c) Capillary pipet from one another) almost simultaneously d) Tissue paper reported three immunologic procedures for the detection of HCG, these procedures were: e) Alcohol and water. f) Місговоре 1. The gel-diffusion method g) Slides 2. A complement-fixation technic h) About 20 cc. of first morning urine or 8 cc. of 3. The hemagglutination-inhibition (HAI) test whole blood if serum is to be used. employing HCG-coated red blood cells. NOTE: Indications of male frogs: Other tests that were also introduced from then on a) Black spots in the neck. includes a radioimmunoassay for HCG, precipitin b) Long, big front claws used in grasping the female test, and the latex agglutination inhibition (LAI) during copulation. technic. The tests most generally adapted in clinical c) Much smaller in size than female. laboratories were the hemagglutimation-inhibition and the latex agglutination-inhibition technics. Procedure: a) Examine the frog before use for presence of The basis of all immunologic tests for pregnancy is spermatozoa. Extract urine from the cloaca by the antigenicity of human chorionic gonadotropin inserting the tip of capillary pipet and place a few (HCG). drops on a glass slide and examine under the low power objective of the microscope. Do not use frogs which are positive for sperm cells. 1. Hemagglutination-inhibitiopn (HAI) Pregnancy Test b) If the frog is negative for sperm cells, then use it for the test. Principle: If urine containing HCG is added to the antiserum con- taining anti-HCG antibody with its c) Inject 2 cc. of urine into the muscle of the thigh antibody (HCG-antiserum), the hormone will and then to the dorsal lymph sae subcutaneously. combine with its antibody and neutralize it. When d) Set the frog aside for 30 minutes then examine the HCG-coated carriers (red blood cells or latex the urine, of the frog for sperm cells. If negative. do par- ticles are then added, no agglutination occurs the next examina- tion after an hour, if still negative, (agglutination is inhibited). This is a positive test. then after 1 hour and a half and so on until 2 hours. a. Gravindex "Slide Test for Pregnancy" e) Positive result is shown by the presence of sperm PROCEDURE: cells in the urine of the frog. Sperm cells are banana shaped. a) Place 1 drop of Gravindex anti-serum on the slide. NOTE: False positive result is obtained during the summer season, since this is their mating period and b) Add 1 drop of urine to be tested using a dropper spermatozoa are present at this time. This can be pipet. avoided however, by tasting the frog's urine before c) Mix well with applicator stick, rock slide gently for using it for the test. 30 seconds. Sensitivity: d) Add 2 drops thoroughly shaken Gravindes Rana pipiens (Male North American Leopard frog) antigen. Mix well with applicator stick, spreading —---------- 20-80 IU/ml. over an area approxi- mately one and one-quarter inches in diameter. Bufo bufo (Male European toad)--------------------------- -----------8-25 IU/ml. e) Rock slide slowly and gently continuously. Bufo Americanus (Male Toad)------------------------------- f) Observe for agglutination not longer than 2 ----------12-45 IU/ml minutes after mixing. Rana clamitans—------------------------------------------------- Interpretations: --------20-30 IU/ml. Negative Test: Agglutination within 2 minutes The sensitivity of male frogs varies seasonally. In Positive Test: No agglutination within 2 minutes. summer, Rana pipiens may be 400% less sensitive Sensitivity: 1 IU/ml. 6|Page Source: Book/notes of Ma’am Tan  LECTURE | MIDTERM Transcribed by: Faulien & Rhed | Organized by: Angela NOTE: All reagents and the urine specimen must be Known positive or negative reacting urines may be brought to room temperature before use. Protein used as controls to verify the activity of the reagent. substances may cause false positive test. Interpretation of Results: Positive is shown by agglutination. Negative is shown by no Other Hemagglutination-Inhibition Tests: agglutination. a) UCG Test Sensitivity: 3-6 IU/ml. b) Pregnosticon Test c) Prepurex Test C. CHEMICAL PREGNANCY TESTS: d) HCG Test (Less accurate and less popular uses chemicals) e) Dence Pregnancy Test 1. Prenandiol Test This is a test for progesterone in the pregnant 2. Latex Agglutination-inhibition (LAI) Pregnancy woman's urine. It is a color reaction test. A few Tests amount of prenan diol reagent is added to urine and a positive test is shown by a deep orange to dark brown color. Negative is amber. 2. Iodine or Lugol's Reaction Test This is to test the presence of epinephrine substance in pregnant urine. A few cc. of iodine solution as added is to urine, the positive reaction is also shown by a deep orange to deep brown color. D. CLINICIAN'S PREGNANCY TESTS: (Tests done by clinicians or physicians only uses drugs.) 1. Prostigmen withdrawal Bleeding Test The woman is injected with prostigmen substance. If not pregnant, she will bleed or menstruate within 2 Procedure with direct answers of agglutination weeks no bleeding if she is pregnant. means a positive result, no agglutination means a 2. Skin Colostrum Test negative result. Colostrum preparation is injected subcutaneously in Procedure: the forearm of the suspected pregnant woman. If 1) Urine the area in- jected does not swell nor show a reddish coloration, the subject is pregnant. If the area swells a) Cool urine to at least room temperature. Filter and a reddish colora- tion appears, the subject is urine using paper provided by the company selling not pregnant. it. 3. Primodos Oral b) Fill capillary tube to mark with filtered urine; expel into circle on slide. A set of two tablets is taken once a day for two con- secutive days. Within 5 days the woman will bleed if c) Add 1 drop of well-shaken Dap test reagent and she is not pregnant: no bleeding occur if she is mix with stirrer by spreading over the whole pregnant. encircled area. 4. Gestex d) Rock slide gently and slowly for 2 minutes and observe for agglutination. One tablet taken orally for a day. Within 4-5 days the woman will bleed if she is not pregnant, no bleeding 2) Serum occurs if she is pregnant. a) Place 1 drop of diluent into circle on slide. b) Fill capillary tube to mark with serum and add to drop of diluent. Mix with stirrer. c) Add 1 drop of well-shaken DAP test reagent and mix with a stirrer by spreading the whole encircled area. d) Rock slide gently and slowly for 2 minutes and ob- serve for agglutination. NOTE: Abnormal sera containing large amounts of macroglobulins should not be used. Neither should sera from patients suspected of having rheumatoid factor. 7|Page Source: Book/notes of Ma’am Tan