CMV is a clinically relevant pathogen in congenitally infected newborns, as well as in immunodepressed (HIV+, transplanted, cancer, or drig-treated) patients. Its timely identification may have diagnostic and prognostic value.Objectives: Validation of a rapid culture (Cx) method (shell vial) detecting the presence of CMV by immunofluorescent antibodies (IFA), compared to conventional Cx and detection by cytopathic effect (CPE) in different biological samples (urine, blood, biopsies, bronchoalveolar lavage(BAL). Evaluation of CMV excretion in immunodepressed and normal subjects using the shell vial method. Methods: A total of 128 clinical samples (sp) received for CMV culture were studied. In addition, CMV Cx were done of urine sp collected from 31 HIV+ and 40 healthy subjects, paired by age, sex and socioeconomic status. For shell vial Cx technique, sp were inoculated onto human foreskin fibroblast (HFF) monolayers grown on converslips in glass vials; afaataer 16 hours, cells were fixed with cold acetone, exposed to monoclonal antibodies specific to the CMV IE-1 protein, and examined by IF. All sp were also inoculated to tubes of HFF and examined for CPE for 28 days.Results Method validation: Out of 87 urine sp, 16 were (+) and 69(-) by IFA and CPE, 2 were (+) by IFA only; out of 22 blood samples, 2 were(+) and 20 (-) by both methods; out of 9 BLA, 2 were (+) and 7 (-) by IFA and CPE; 10 others sp were (-) by both IFA andCPE. Conventional Cx showed CPE between days 3 and 27 (mean 10.2 days). Population evaluation: 5/31 (16.2%) of HIV+ subjects (mean age 34 years) excreted CMV in the urine; in two of them CMV retinitis was detected following CMV culture result; 3 remain asymptomatic. CMV was not isolated from 40 healthy subjects (mean age34.2 years). DISCUSSION. The shell vial technique compares favourably to conventional Cx, allowing a 24 hr turnaround without decreasing sensitivity. The shell vial assay is particularly useful for immunodepressed subjects who benefit from early diagnosis of infection providing the opportunity for institution of antiviral therapy. CMV infection is frequent among HIV+ patients, and its detection allows for the diagnosis of unnoticed complications.
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(Grant FONDECYT 92-0656).
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Vial, P., Ferrés, M., Ovalle, J. et al. Immunodetection of Cytomegalovirus (CMVO Immediate Early (IE-1) Protein in Tissue Culture at 16 Hours Incubation: Validation and Comparison to Conventional Culture, and Evaluation of Normal and Immunodepressed Pol. Pediatr Res 39, 371 (1996). https://doi.org/10.1203/00006450-199602000-00050
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DOI: https://doi.org/10.1203/00006450-199602000-00050