[Complete genome sequencing of enterovirus 71 (EV71) HENAN08 strain isolated in Henan province].

OBJECTIVE: To sequence the whole-genome of enterovirus 71 (EV71) srtain isolated from patient with hand, foot and mouth in Henan province in 2008. METHODS: Eight overlapping clones covering the whole viral genome were obtained by RT-PCR and the sequences were determined by Sanger dideoxg-mediated chain termination method. RESULTS: Data it showed that the full length of enterovirus 71 (EV71) HENAN08 genome (not including Poly A tail) is 7405 bp. No deletion or insertion was detected in the coding region. There were several deletions and insertions in 5'UTR and 3'UTR regions. In P1 region, HENAN08 strain shared high homology with AnhuiFY08 strain, Zhejiang08 strain and SHZH strains (SHZH98, SHZH03) but low homology with Cox. A16. In P2 and P3 regions, HENAN08 strain shared higher nucleotide homology with Cox. A16 (81.7% and 83.7%) than that with BrCr and TW2086 strains. The phylogenetic analysis based on P1 region demonstrates that HENAN08 strain had the nearest genetic relationship with AnhuiFY and Zhejiang strains (isolated in 2008). CONCLUSION: The HENAN08 strain might belong to the same genogroup with AnhuiFY08 and Zhejiang08 strains as C4 gene subtypes.

[Construction for detecting of human papillomaviruses types by using a multiplexed Luminex assay].

AIM: To develop multiplex human papillomavirus genotyping, a quick and sensitive high-throughput procedure for the identification of multiple HPV genotypes by using a multiplexed Luminex array. METHODS: Standard plasmids containing HPV genomic DNA were developed a standard multiplexed suspension array for detection 13 HPV types on Luminex analysis platform. the primer and probes were selected, HPV type-specific oligonucleotide probes were coupled to fluorescence-labeled polystyrene beads. RESULTS: The one or two of mixed HPV standard plasmids were identified HPV type by using 13 HPV genetypes multiplexed suspension array, different HPV types can be correctly detected simultaneously. The positives of HPV clinical samples were detected by using this method, and the genetypes of samples by detecting were the same results with using BLAST in NCBI. CONCLUSION: The proposed method allowed a high through-out, special, simple, rapid and economical identification of HPV DNA genotypes, It is expected to be an extremely useful tool in the identification of HPV genetypes for a lot of clinical samples.