Genotyping Of Congenital Cytomegalovirus Infection In HeFei

Objective Normal maternal blood were isolated and identified HCMV. To assess the congenital cytomegalovirus infection and investigate the gB genotypes of cytomegalovirus circulating in HeFei, China. In order to understand the features of HCMV molecular epidemiology in HeFei . To provide the basis for the further study the pathogenicity, immunity and the development of HCMV subunit vaccine.Methods We collected 160 cases of umbilical cord blood samplesfrom the normal maternal women (aged 19 to 38 years) in Maternal and Child Health Hospital, HeFei, Anhui. Take the samples to test : (1) laboratory diagnosis of congenital cytomegalovirus infection: HCMV isolation, observing cytopathic effect (CPE) of HCMV; HCMV pp65 antigen by indirect immunofluorescence assay (IFA),HCMV DNA IE and UL83 polymerase chain reaction (PCR) and HCMV pp65 antigenemia assay .These methods were all used to assess the rate of congenital cytomegalovirus infection . (2) genotyping of congenital cytomegalovirus infection: each positive samples were genotyped by ristriction fragment of length polymorphism (RFLP) with Rsa I and Hinf I. These strains were carried out gB gene PCR amplification, sequenced, aligned and analysised with strain of HCMV isolates on Genbank .Drawn phylogenetic tree by the MEGA 4.0 software . (3) We do retrospective study and further track the clinical outcomes of children with congenital cytomegalovirus infection.Results (1) laboratory diagnosis of congenital cytomegalovirus infection and diagnostic criteria of congenital cytomegalovirus infection : viral isolation rate was 25.0%(40/160); Positive rate of HCMV pp65 IFA was 24.4%(39/160);.These methods were assessed congenital cytomegalovirus infection. We get that the rate of congenital cytomegalovirus infection in HeFei is 25% after analysis. (2) genotyping of congenital cytomegalovirus infection: the positive samples are genetyped by RFLP. They were all determined to be gB-2 type. It can be seen that congenital cytomegalovirus infection in the region is all gB-2 type. Genotypes have also found that HeFei-01, HeFei-03 and HeFei-21 samples have 24 (7.95%) mutations, 28 (9.27%) and 30 mutations (9.93%).These results suggeste that these virus strains have a higher frequency of recombinantion. (3)So we do retrospective study and track the long-term clinical outcomes of children with congenital cytomegalovirus infection especially samples of HeFei-01, HeFei-03 and HeFei-21.Conclusion (1) The rate of newborns with congenital cytomegalovirus infection was 25.0%;(2) HCMV gB gene sequence isolates and phylogenetic tree analysis showed that : We named the positive samples as HeFei-01 to HeFei-40.The data from this study showed congenital cytomegalovirus infection in HeFei were all gB-2 type. (3) However, additional genetic variations in gB gene exist and it could be explained as a result of homologous recombination. Samples HeFei-01, HeFei-03 and HeFei-21 have a higher frequency of recombinant. (4) Birth weight of HeFei-21 Newborns is only 2.2 kg which meets the diagnostic criteria of intrauterine growth retardation. It is necessary to establish a follow-up file and do medical examination to these 40 cases of congenital medical examination infection.