Clinical experience with cytomegalovirus isolation using conventional cell cultures and early antigen detection in centrifugation-enhanced shell vial cultures.
A total of 1,915 clinical samples was inoculated by low-speed centrifugation into shell vials (Bartels Immunodiagnostics, Bellvue, Wash.) containing cover slip monolayers of MRC-5 fibroblasts. At 1 and 2 days postinoculation, one cover slip was stained by an indirect immunofluorescence technique using a monoclonal antibody (Biotech Research Laboratories for Dupont, Billerica, Mass.) to cytomegalovirus (CMV) early antigen (EA). Clinical samples were also inoculated into three MRC-5 or MRHF cell cultures which were observed for 30 days for the appearance of a cytopathic effect (CPE). Of 157 CMV-positive samples, 92 (59%) were identified by centrifugation-enhanced EA (CE-EA) and 131 (83%) produced a CPE. CE-EA was less sensitive than CPE for all types of samples, although 17% of CMV-positive samples were detected by CE-EA alone. Evaluation of the CMV status of patients with CE-EA-positive-CPE-negative samples indicated that these samples likely represented true CMV-positive results. The average elapsed time between culture inoculation and identification of CMV decreased as follows when both CE-EA and CPE, rather than CPE alone, were used: urines, 15 to 7 days; buffy coats, 18 to 9 days; lung samples, 13 to 8 days; throat samples, 18 to 7 days. Although CE-EA was less sensitive than 30-day cell culture, both CE-EA and CPE were identified as valuable in CMV detection, and neither could be discontinued without a decrease in the CMV isolation rate or an increase in the turnaround time.
Leland
D S
DS
Department of Pathology, Indiana University Medical Center, Riley Hospital, Indianapolis 46223.
Hansing
R L
RL
French
M L
ML
D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis*; D002460:Cell Line; D002498:Centrifugation; D003587:Cytomegalovirus / Q000276:immunology / Q000302:isolation & purification*; D003588:Cytopathogenic Effect, Viral; D005347:Fibroblasts; D005455:Fluorescent Antibody Technique; D006801:Humans; D017874:Immediate-Early Proteins; D008168:Lung / Q000382:microbiology; D010614:Pharynx / Q000382:microbiology; D011237:Predictive Value of Tests; D013997:Time Factors; D014556:Urine / Q000382:microbiology
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D017874:Immediate-Early Proteins; C044582:immediate-early proteins, cytomegalovirus