Objective To investigate the mutations in nueleotide and amino acid level in HPV61,83 and 84 Shanxi isolates.Methods Amplified fragments of HPV61,83 and 84 from human papillomavirus (Human papillomavirus,HPV) molecular epidemiologic survey of Shanxi Province using HPV consensus primers MY09/11 were cloned in pMD18-T vector,and the plasmids were sequenced,then nucleotide sequences and amino acid sequences were analyzed.Results HPV61 and HPV83 isolates were consistent with reference strains U31793 and AF151983 in nucleotide sequences;four mutations of nucleotide (C6760T,T6931C,T6951C and C6987A) were found in HPV84 isolate compared with reference strain AF293960,among which C6987A resulted in D441E and the amplified sensitivity of standard sample of HPV61 using primers MY09/11 was higher than that of HPV83 and 84.Conclusion HPV61 and HPV83 isolates were consistent with reference strains,four mutations of nucleotide and one mutation of amino acid were found in HPV84,the amplified sensitivity of standard sample of HPV61 using primers MY09/11 was the highest among those three isolates.%目的 探讨山西省的HPV61、83和84型分离株MY片段中核苷酸及氨基酸的突变情况以及山西省分离株与α3中其他型别的同源性,制备三型别包含MY片段的标准品.方法 使用HPV通用引物MY09/11对我国HPV61、83和84型分离株进行PCR扩增,并连接至pMD18-T载体,所得质粒进行序列分析,应用BioEdit生物学软件进行核苷酸水平和氨基酸水平的分析.结果 HPV61和83型分离株分别与参考株序列U31793和AFl51983比对,在核苷酸水平完全一致;HPV84型分离株与参考株序列AF293960比对,在核苷酸水平发生4个点突变,分别为C6760T、T6931C、T6951C和C6987A,其中C6987A导致D441E氨基酸残基的突变;MY-PCR反应对HPV61、84和83型标准品检测的灵敏度依次降低.结论 我国HPV61和83型分离株与参考株序列相比较未发现突变,HPV84型与标准株相比存在着核酸和氨基酸水平的突变;MY09/11通用引物检测HPV61型标准品的灵敏度高于HPV83和84型.
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