Correlation of cytomegalovirus (CMV) detection in urine by tissue culture virus isolation, early antigen immunofluorescence test and nucleic acid hybridization

Abstract

The sensitivity and specificity of the early antigen immunofluorescence test and of the hybridization assay were compared with those of the development of a cytopathic effect on HF MA 23 cell monolayers in tissue culture. Of 818 clinical specimens tested, 125 specimens produced CMV cytopathic effect in tissue culture within four weeks after inoculation. Of the 125 specimens, 71 were positive for CMV after 36 h by early antigen immunofluorescence test (sensitivity 56.8%). Of the remaining 693 culture-negative specimens, seven were positive by early antigen immunofluorescence test (specificity, 99%). A hybridization assay using a commercial kit with the biotinylated Bam H I AB DNA fragment of HCMV Towne strain and horseradish peroxidase streptavidin conjugate, detected 60 pg of HCMV DNA. In this assay 104 urine specimens were tested. Compared with tissue culture, this hybridization assay had a sensitivity of 69.5%, a specificity of 60.3% and a correlation of 63.4%, at which 25 of 33 tissue culture positive samples had positive hybridization results, and 27 of 68 tissue culture negative samples also gave positive hybridization results. An additional hybridization assay, using the biotinylated Hind III L fragment of HCMV AD 169 and 125J streptavidin conjugate, detected 30 pg HCMV DNA. In 179 urine specimens simultaneously tested by tissue culture, 28 of 33 positive tissue culture specimens were positive in the hybridization assay. 37 of 146 negative tissue culture specimens gave positive hybridization signals (sensitivity 84.8%, specificity 74.7%, correlation 76.5%).