Multicenter Comparison of the Digene Hybrid Capture CMV DNA Assay (Version 2.0), the pp65 Antigenemia Assay, and Cell Culture for Detection of Cytomegalovirus Viremia

Abstract

We compared the Digene Hybrid Capture CMV DNA Assay version 2.0, the pp65 antigenemia assay, traditional tube culture, and shell vial culture for the detection of cytomegalovirus (CMV) viremia in several patient populations at three centers. Of 561 blood specimens collected from 402 patients, complete clinical and laboratory data were available for 489. Using consensus definitions for true positives and true negatives, the sensitivities of the Hybrid Capture assay, antigenemia, shell vial, and tube culture were 95, 94, 43, and 46%, respectively. The specificities of the Hybrid Capture assay and antigenemia were 95 and 94%, respectively. At all three study sites, the detected level of CMV viremia was significantly higher with the Hybrid Capture assay or antigenemia than with shell vial and tube culture. In a group of 131 healthy nonimmunosuppressed volunteers, the Hybrid Capture assay demonstrated a specificity of over 99%. The Hybrid Capture assay is a standardized assay that is simple to perform and can utilize whole blood specimens that have been stored for up to 48 h. The high sensitivity and specificity of the Hybrid Capture assay along with its simplicity and flexibility make it a clinically useful assay for the detection of CMV viremia in immunocompromised or immunosuppressed patients. Further evaluation to determine its role in predicting CMV disease and for monitoring the therapeutic response to anti-CMV therapy is needed.

Role of Cell Culture for Virus Detection in the Age of Technology

“…Within the past decade, IF methods, called CMV antigenemia assays, have been applied for direct detection and quantification of CMV antigens in peripheral blood leukocytes (65,103,104). A thorough review of CMV antigenemia methods and clinical applications has been published previously (10).…”

“…The original methods used HRP stains, but FITC quickly replaced HRP. CMV detection in 3 to 4 h via antigenemia assays was a major advance in dealing with detection of CMV, which can require 10 to 30 days to produce CPE in traditional tube cell cultures (65,103,104). An added advantage is the semiquantitative numerical value (number of fluorescent cells per number of leukocytes per slide) obtained.…”

“…Sensitive and rapid CMV detection in shell vials was seen in specimens from most body sites; however, of concern was the 25% of positive blood samples detected by traditional tube culture alone (120). This study and others have prompted investigators to suggest that if culture alone is to be used, both shell vial and conventional tube culture systems must be used for optimal CMV recovery (103,104,120).…”

Infection in Renal Transplant Recipients

“…Quantitation of the intensity of CMV infection has been linked to the risk for infection in transplant recipients. 7,33,42,50,65 Two types of quantitative assays have been developed-molecular and antigen detection assays. The antigenemia assay is a semiquantitative fluorescent assay in which circulating neutrophils are stained for CMV early antigen (pp65) that is taken up nonspecifically as a measure of the total viral burden in the body.…”

Application of Molecular Diagnostic Techniques for Viral Testing

“…The antibody conjugate is detected with a chemiluminescent substrate and a luminometer serves as a device to measure the light emitted. Hybrid capture assays for detection of human papillomavirus (HPV) and cytomegalovirus (CMV) in clinical specimens are currently commercially available (Digene Corp., Gaithersburg, MD) [12,13].…”