Epidemiological studies of HCMV infection in children by restriction analysis of long-PCR products

摘要

Sequences of MIE, pol, gB gene longer fragments (2.0-2.6 kb) were amplified in two stage PCR (nested PCR) from 46 PEG-urine extracts. Restriction of particular amplicons with selected endonucleases identified 19 distinct HCMV genotypes: 8 for MIE, 6 for gB and 5 for poi. MIE profiling revealed highest differentiation power. Our data showed the correlation among MIE, pol, and gB genotypes and clinical disease, indicating that genotypes may contribute the course of HCMV infection. For newborns with symptomatic infection, the highest level of variability among three genes analysed was obtained in comparison to samples of newborns with asymptomatic infection. Long PCR technique coupled with restriction analysis may be successfully applied in HCMV genotypes differentiation in samples taken directly from HCMV infected individuals, thus may be useful in prognosis of clinical infection.