| Torque teno virus is a nonenveloped virus with a small, covalently closed circular genome of single-stranded DNA. TT virus is a relatively new virus obtained in1997by differential display technology from a hepatitis patient without any know virus marker for know hepatitis virus. The study found a variety of TT virus can infect people and animals, and a high prevalence in the population.TT virus was distributed worldwide. It has a high degree of genomic variability in human and animal epidemic situation is not the same, and the current lack of domestic research in different areas of human TT virus more epidemiological research data, which include virus the genotype and infection. In addition, both in vivo and in vitro experiments, the research progress of TT virus is relatively slow, especially the virus in the epidemiology of healthy people. Therefore, in order to understand the molecular epidemiology of the virus, and genome characteristics, as well as Torque teno viral disease that may bring prevention to provide a scientific basis for the experimental serum samples of healthy people in Kunming launched a TT virus molecular epidemiology of infection.In this study, the extraction of Kunming healthy human serum viral genome, according to the GenBank sequence published TTV standard strain (AB008394), and with reference to the relevant literature, design and synthesis of the nucleotide sequence for the more conservative TT virus untranslated region (Untranslated Region, UTR) of primers were used in a nested PCR assay for detection of human TT virus. Blood samples of224copies of a university in Kunming healthy people were tested, the results showed that healthy people TT virus infection rate reached67.41%. According to the research have been published abroad, for Torque teno viral nucleotide sequence variation rate is relatively large open reading frame1(Open Reading Frame one, ORF1) were designed and synthesized primers N22area, while the fourth was synthesized TT virus genes (Group4, G4) and the fifth gene cluster (Group5, G5) genotyping primers to amplify a good step for the sample of120were classified experiments. Identified by PCR primers for screening were genotyped for the serum samples positive. Which is better for the PCR amplification of samples for sequencing and analysis, phylogenetic tree. Typing results showed, N22district primer21, accounting for17.5%of the experimental samples to G4primer50sample,13were positive detected, the ratio of26%, G5primers50copies,24copies detected, the positive rate was48%. Most strains with TTV1a type match, but a little distant evolutionary relationship may be caused by a mutation of the gene. Designed and synthesized five pairs of primers for TT virus genome were cloned and sequenced, and the sequence assembly, the establishment of this strain of the virus phylogenetic tree analysis of amino acid homology. Phylogenetic analysis showed that this strain belongs TTV1a type. The nucleotide and amino acid sequence homology analysis showed the highest similarity with TTV1a type.The above findings suggest that TT virus in healthy people Kunming detection rate of67.41%in Kunming infer that this virus still has a higher prevalence; genotyping and phylogenetic analysis showed that the advantages of the popular Kunming or TT virus strains of type1a.In this study, healthy population of Kunming molecular epidemiology of Torque teno virus made a more systematic investigation, the results of the above study was TT virus in healthy population prevalence and genotypes provide a reference for TT virus further research has laid a solid foundation. |
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