Lab Dept:

Microbiology/Virology

Test Name:

CMV RAPID FA

General Information

Lab Order Codes:

RCMV

Synonyms:

CMV Shell vial culture; Cytomegalovirus Rapid FA

CPT Codes:

87254 x2 - Virus isolation; shell vial, includes identification with immunofluorescence stain, each virus

Test Includes:

Shell vial isolation technique with immunofluorescent staining of CMV early nuclear antigen. Viral culture must be ordered with this test. Refer to Viral Culture

Logistics

Lab Testing Sections:

Virology

Phone Numbers:

Minneapolis:

Saint Paul:

612-813-5806

651-220-6555

Test Availability:

Daily, 24 hours

Turnaround Time:

1 - 2 days

Special Instructions:

Do Not use calcium alginate swabs.

Requisition must state specific site of specimen and date/time of collection.

Specimen

Specimen Type:

Urine, throat, bronchoalveolar lavage, bronchial washings, appropriate autopsy and biopsy specimens, blood, bone marrow, tracheal aspirates

Container:

Viral transport media (M4 VTM) (available in Microbiology), sterile container, swab transport system, Lavender top (EDTA) tube

Volume:

Whole blood: 5.0 mL

Urine: 5.0 mL

Washings/aspirates: 1.0 - 2.0 mL

1 swab

Aspirate or sputum: 0.5 mL

Collection:

Blood:

Venipuncture for patients less than 2 months of age: Prep with 2% tincture of iodine:

1. Disinfect the stopper of the Lavender top (EDTA) tube with 70% alcohol. Allow to dry.

2. Scrub venipuncture site with 70% alcohol for 1 min using the Frepp® applicator. Allow to dry.

3. Using the Sepp® applicator, apply 2% tincture of iodine to site starting at the center and moving outward in concentric circles. Allow to dry.

4. If the site must be touched during venipuncture, disinfect the gloved fingers.

5. Collect 5.0 mL of blood and aseptically inoculate the Lavender top (EDTA) tube.

6. Gently invert the tube 4-5 times to mix contents.

7. Following collection, remove the iodine using the Frepp® applicator or an alcohol pad.

Venipuncture for patients more than 2 months of age: Prep with CloraPrep Sepp® Applicator.

1. Disinfect the stopper of the Lavender top (EDTA) tube with 70% alcohol and allow to dry.

2. Break the Sepp® ampule to release the 2% CHG.

3. Apply the CloraPrep solution using a back-and-forth friction scrub for 30 seconds.

4. Allow the area to dry for 30 seconds.

5. If the site must be touched during venipuncture, disinfect the gloved fingers.

6. Collect 5.0 mL of blood and aseptically inoculate the Lavender top (EDTA) tube.

7. Gently invert the tube 4-5 times to mix contents.

Line Draw:

1. Prep catheter port with 2% tincture of iodine or betadine followed by 70% alcohol. Allow to dry.

2. Aseptically collect 5.0 mL of blood through the injection port. Blood may be collected without first drawing a discard.

3. Aseptically inoculate the Lavender top (EDTA) tube.

Do Not centrifuge. Send in original Vacutainer^™ tube. Forward unprocessed whole blood promptly at ambient temperature only.

Bone Marrow:

Place 1.0 – 5.0 mL of bone marrow in Lavender top (EDTA) tube(s). Invert several times to mix bone marrow. Do Not centrifuge. Send in original Vacutainer^™ tube. Forward unprocessed bone marrow promptly at ambient temperature only.

Throat Swab:

1. Depress the tongue with a tongue depressor so the swab does not touch the tongue.

2. Sample the posterior pharynx, tonsils, and inflamed areas with a sterile swab.

Tissue:

Submit specimen in a screw capped, sterile container.

Urine:

Males:

1. Clean glans with soap and water.

2. Rinse area with wet gauze pads.

3. While holding foreskin retracted, begin voiding.

4. After several mL have passed, collect a minimum of 5.0 mL without stopping flow of urine.

5. Maintain sterility and forward immediately to the Microbiology Lab. Refrigerate.

Females:

1. Thoroughly clean urethral area with soap and water.

2. Rinse area with wet gauze pads.

3. While holding labia apart, begin voiding.

4. After several mL have passed, collect a minimum of 5.0 mL without stopping flow of urine.

5. Maintain sterility and forward immediately to the Microbiology Lab. Refrigerate.

Bronchoscopy:

1.0 – 2.0 mL of specimen obtained by physician through the biopsy channel of the bronchoscope.

Transfer specimen into a luki tube. Transport to the Microbiology Laboratory immediately.

Special Processing:

Extract swabs into viral transport media (VTM) by swirling and pressing the swab against the inside of the vial, then discard swab; add 3.0 - 5.0 mL urine to urine VTM; place washings/aspirates into VTM; place tissue into VTM. Refrigerate.

Transport/Storage:

Maintain sterility and transport to the Microbiology Lab immediately at room temperature. Store at refrigerated temperatures.

Note: If there is a delay in transport of 1 hour or more, place specimen in viral transport media and refrigerate.

Sample Rejection:

Specimen with a transit time exceeding 2 hours after collection; specimen not submitted in appropriate transport container; improperly labeled specimen; insufficient volume; external contamination. If an unacceptable specimen is received, the physician or nursing station will be notified and another specimen will be requested before the specimen is discarded.

Interpretive

Reference Range:

No Cytomegalovirus detected by rapid fluorescent antibody.

Critical Values:

Positive results in systemic infections will be called to the physician or nursing unit.

Limitations:

● Urine and blood can be toxic to cell cultures and can result in inconclusive results.

● CMV antigenemia or molecular techniques are more sensitive tests for the detection of CMV in blood.

Methodology:

Shell vial culture with immunofluorescence

Additional Information:

● CMV infections are very common in normal individuals and are usually asymptomatic. However, CMV infections are frequently severe and life threatening in immunocompromised patients, including organ recipients and AIDS patients. CMV is the major viral pathogen following renal transplantation. Blood cultures positive for CMV predict progression. Knowledge of CMV infection is of utmost importance so that ganciclovir can be started as soon as possible.

● CMV is the most frequent cause of congenital viral infections in humans and occurs in about 1% of all newborns. Approximately 90% have no clinical symptoms at birth. Ten percent to 20 % of these infants will develop complications before school age. Congenital infection may occur as a result of either primary or recurrent maternal infection.

References:

Cook, JH, and M Pezzlo (1992). Specimen receipt and accessioning. Section 1. Aerobic bacteriology, 1.2.1-4. In HD Isenberg (ed) Clinical Microbiology Procedures Handbook. American Society for Microbiology, Washington DC

Miller, J Michael (1999) A Guide To Specimen Management in Clinical Microbiology, American Society for Microbiology, Washington DC

Miller, J Michael, and HT Holmes (1999) Specimen Collection, Transport, and Storage In PR Murray et al, (ed), Manual of Clinical Microbiology, 7^th edition, American Society for Microbiology, Washington DC, pg 33-104

Griffiths, PD, and VC Emery (2002). Cytomegalovirus In DD Richman et al., (ed.), Clinical Virology, 2nd edition, American Society for Microbiology, Washington DC, pg 447-449