CYTOMEGALOVIRUS is most prevalent among very young children. Major areas of risk include prenatal or postnatal infants and immunocompromised individuals. HCMV is considered an AIDSdefining infection In HIV infected persons.
CYTOMEGALOVIRUS is most prevalent among very young children. Major areas of risk include prenatal or postnatal infants and immunocompromised individuals. HCMV is considered an AIDSdefining infection In HIV infected persons.
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CYTOMEGALOVIRUS is most prevalent among very young children. Major areas of risk include prenatal or postnatal infants and immunocompromised individuals. HCMV is considered an AIDSdefining infection In HIV infected persons.
Copyright:
Attribution Non-Commercial (BY-NC)
Available Formats
Download as PPTX, PDF, TXT or read online from Scribd
particularly those live in crowded conditions. ?
oral, respiratory or venereal routes, through organ transplant, or fresh transfused blood. Without any detectable signs or symptoms. ? pome of them develop a fever-like syndrome, with prolonged fever, and a mild hepatitis. ? ëajor areas of risk of infection include pre- natal or postnatal infants and immunocompromised individuals, such as organ transplant recipients, persons with leukemia, or those infected with human immunodeficiency virus (HIV). In HIV infected persons, HCëV is considered an Õ
, indicating that the T-cell count has dropped to low levels.
? Yrine, saliva, blood and biopsy samples can be used for virus isolation. ? Yrine should be collected a sterile container without additives. ? paliva samples should first be soaked on to a swab which is then broken off into transport medium. ? Blood should be collected into a heparinized bottle (some phenolic preservatives found in proprietary pathology blood bottles may be toxic to blood cultures) containing 500 units of heparin. ? Tissue biopsies should be placed in sterile plastic containers. The specimens can be treated in the following ways p IC T pT F CëV ? Passive latex agglutination @ ëethod for the detection of antibodies to CëV involves the mixing of latex particles with the patient·s serum.. ? ë TH 1 : PAppIV AT AYTINATIN F TH T CTIN F ANTIBI p T CYTë AVIYp IN HYëAN p Yë ? ëAT IAp ? The kit contains : 1. CëV antigen-coated latex participles prepared from disrupted CëV that has been judged to be inactivated by bioassay procedures. 2. Phosphate-buffered saline , PH 7.4, containing bovine serum albumin with 0.02 percent sodium azide. 3. Test cards, which must be flat for proper reactions . 4. Plastic stirrers 5. ispensing needle , 2-1 gauge, green hub . 6. High-reactive control serum (human) with 0.1 percent sodium azide . 7. ow-reactive control-serum (human) with 0.1 percent sodium azide . 8. Nonreactive control serum (human) with 0.1 percent sodium azide . ? Additonal material needed (not provided by the kit ): 1. Centrifuge 2. otator with humidifying cover . 3. High-intensity incandescent lamp . 4. ëicropipettors, 25 microliter delivery . 5. Vortex mixer . pP CIë N YI ë NTp ? A minimum of 2 ml of clotted blood or anticolagulated promptly and an aliquot of serum or plasma removed plasma specimens containing TA or heparin as an anticoagulant can be used for qualitative or quantitative testing using the same technique as for serum samples . ? ppecimens may be stored for up to 1 week in the refrigerator.18 degrees C if longer freezing PC Y (YAITATIV ) ? Before beginning the procedure , allow the reagents to reach the room temperature . do not mix reagents to different kit lot of numbers and avoid microbial contamination of reagents . label each circle of the card with appropriate identification of patient sera and controls . 1. Ysing a micropipettor , place 25 microliter of each specimen 2. Ysing a new plastic stirrer for each circle , spread the serum to fill the entire circle . 3. Hold the bottle cap over the tip of the needle and gently invert the latex reagent dispensing the bottle several times. 4. ispense 1 free-falling drop (approximately 15 microliter ) 5. Hand rotate the card back and forth 3 or 4 times to distribute the latex antigen throughout the circle. 6. Place the card on the rotator and mix for 8 mins. Ynder a moistened humidify cover. 7. Immediately after rotation, read the card macroscopically in the wet state. Test should be read under a high intensity incandescent lamp. INT P TATIN ? Any agglutination of the latex reagent is regarded as a positive. If the suspension remains evenly dispersed with no agglutination, report as negative. IpCYppIN ? Antibodies may not be detectable in early stage. ? The presence of CëV antibodies in qualitative testing on a single acute-phase is a n indication of previous exposure to the virus but does not indicate immunity to subsequent reinfection. TH FF pHY B NT WITH TH pP CT T THIp PC Y THAT INVV CëV ANTIBY T CTIN:
1. Pt. with acute infection may not be have detectable
antibody. 2. peroconversion may indicate recent infection, but an increase in antibody titer by this method does not differentiate bet. A primary and secondary antibody response. 3. The timing of antibody response during a primary infection may differ slightly. ? The Cytomegalovirus Igë Assay ? This test, which is an indirect enzyme-labeled immunosorbent assay, detects Igë antivodies to CëV, using antigen-coated microwells as a solid phase. Igë antbodies ma persist as long as 9 mos. In immunocompetent pt. ? Ibë responses vary bet. iff indivuals. For example, 10-30% of infants cond=genitally infected with CëV fail to develop Igë antibody ? ë TH 2: CYTë AVIYp Igë AppAY (YANTITATIV )
? The technique describe here is intended for use
with the kit provided by pigma chemical Co. pt ouis, ë ëAT IAp 1. ëicroplate walls coated with CëV antigen. ptore at 2-6 degrees C with desiccant in the reusable plastic bag bag and reseal after opening. 2. Holder or wells 3. iluent, a buffered protein solution,containing surfactant and blue dye,pH 7.5 4. Calibrator. This is the human serum containing Igë antibodies to CëV at 100 arbitrary units. 5. Conjugate. Contains goat antibodies to human Igë labeled with calf alkaline phosphatase. 6. pubstrate. This contains p-nitrophenyl phosphate, disodium, and hexahydrate 1mg/ml, pH 9.6 . 7. Wash concentrate. This is buffer solution concentrate with surfactant. 8. ptop solution. An alkaline solution pH 12.0. ptore at room temp. 9. Positive control. This is human serum containing Igë antibodies to CëV and .1 percent sodium azide as a preservative. 10. Negative control. This is human serum containing no detectable antibodies to CëV and .1 percent sodium azide as a preservative. AITINA ëAT IA YI 1. ppectrophotmeter 2. Pipettes 3. Timer 4. 1 liter measuring cylinder 5. pqueeze bottle for dispensing wash solution 6. ilution plates 7. Test tubes or cuvettes pP CIë N YI ë NT ? ike the first method. ? If the test cannot be performed immediately, the specimen should be refrigerated or frozen. PC Y 1. ilute callibrator, positive and negative controls and test samples by combining 10 micro liter of each with 200 micro liter of sample diluent in tubes or diluent plates. 2. Placed the desired no. of antigen wells in the holder. 3. Ysing a pipette tip, mix the samples and the diluent by drawing up and expelling two or three times. 4. Include one well that contains only 100 micro liter sample diluent. 5. Allow the plate to stand at room temp for 30 (+- 2 ) mins. 6. phake out or aspirate the contents of the wells. 7.Place 2 drops (100 Y) conjugate in each well, including the reagent blank well. 8. Allow to stand at room temp 3o mins. 9. Wash wells by repeating step 6. 10. Place 2 drops (100 Y) substrate in each well, including the reagent blank well. 11. Allow to stand at room temp 3o mins. 12. Place 2 drops (100 Y) stop solution in each well, including the reagent blank well. 13. ead and record absorbance of each test at 405 nm within 2 hours after the reaction has been stopped. IpCYppIN ? False negative results can be due to a low level of specific Igë antibodies or interference by competitive antigen specific CëV-specific Ig( specially in congenitally infected newborns because of the presence of maternal Ig) ? False positive results can be due to interference by Igë rheumatoid factor.