人乳头瘤病毒基因分型液态芯片的构建

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

目的:通过构建人乳头瘤病毒基因分型的液态芯片,建立高通量、灵敏和特异的HPV型别检测系统。方法:在LuminexTM平台上,采用国际公认的标准HPV质粒建立13种基因分型HPV芯片,选择引物及探针,将探针有效的偶联到微球上。结果:用LuminexTM平台建立了13型HPV芯片,对单一或两种标准质粒混合的样品均可准确分型。对分型检测的阳性标本进行基因测序,经美国NCBI数据库BLAST比对符合其检测型别。结论:利用LuminexTM平台构建了13种HPV型别液相芯片的诊断体系,具有高通量、快速准确等特点,适用于大规模临床样本的HPV基因分型的诊断。

AIM： To develope 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^TManalysis 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 identificated HPV type by using 13 HPV genetypes multiplexed suspension array, different HPV types can be correctly detected simultaneously. The postives 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-hout, 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 clincial samples.