Analysis of cytomegalovirus gene by isolating fetal cells in maternal blood

Abstract

To establish a new criteria of noninvasive prenatal diagnosis of human cytomegalovirus intrauterine infection by isolating single fetal cell in maternal peripheral blood. Micromanipulation techniques were applied to isolate single fetal nucleated erythroblasts from 273 maternal blood samples. SRY gene and HCMV DNA of the single fetal cell were detected by multiple primed in situ labeling (PRINS) from 76 samples of maternal peripheral blood which were HCMV-DNA positive. SRY gene and HCMV DNA of a single fetal cell were detected by primed extension preamplification (PEP) and polymerase chain reaction (PCR) from 273 samples of maternal peripheral blood. The detection rate of fetal cells from maternal blood was 100% by micromanipulation techniques. The sensitivity of detecting SRY gene by PRINS was 97. 56% (40/41 ) and its specificity was 100% (35/35). The sensitivity and specificity of detecting SRY gene by PEP and PCR were 97.39% (149/153) and 99.17% (119/120) respectively. The sensitivity and specificity of detecting HCMV-DNA by PEP and PCR were 95. 12% (39/41) and 100% (232/232) respectively. The new method of noninvasive prenatal diagnosis of human cytomegalovirus intrautefine infection by isolating single fetal cell in maternal peripheral blood by use of PRINS, PEP and PCR was of high sensitivity and specificity and may be widely used in clinical laboratory.