中华实验和临床病毒学杂志
中華實驗和臨床病毒學雜誌
중화실험화림상병독학잡지
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
2010年
2期
107-109
,共3页
王宝晨%王晓莉%薛凤霞%刘宏图
王寶晨%王曉莉%薛鳳霞%劉宏圖
왕보신%왕효리%설봉하%류굉도
乳头状瘤病毒,人%标准品%基因型
乳頭狀瘤病毒,人%標準品%基因型
유두상류병독,인%표준품%기인형
Papillomavirus,human%Standard sample%Genotype
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.
目的 探討山西省的HPV61、83和84型分離株MY片段中覈苷痠及氨基痠的突變情況以及山西省分離株與α3中其他型彆的同源性,製備三型彆包含MY片段的標準品.方法 使用HPV通用引物MY09/11對我國HPV61、83和84型分離株進行PCR擴增,併連接至pMD18-T載體,所得質粒進行序列分析,應用BioEdit生物學軟件進行覈苷痠水平和氨基痠水平的分析.結果 HPV61和83型分離株分彆與參攷株序列U31793和AFl51983比對,在覈苷痠水平完全一緻;HPV84型分離株與參攷株序列AF293960比對,在覈苷痠水平髮生4箇點突變,分彆為C6760T、T6931C、T6951C和C6987A,其中C6987A導緻D441E氨基痠殘基的突變;MY-PCR反應對HPV61、84和83型標準品檢測的靈敏度依次降低.結論 我國HPV61和83型分離株與參攷株序列相比較未髮現突變,HPV84型與標準株相比存在著覈痠和氨基痠水平的突變;MY09/11通用引物檢測HPV61型標準品的靈敏度高于HPV83和84型.
목적 탐토산서성적HPV61、83화84형분리주MY편단중핵감산급안기산적돌변정황이급산서성분리주여α3중기타형별적동원성,제비삼형별포함MY편단적표준품.방법 사용HPV통용인물MY09/11대아국HPV61、83화84형분리주진행PCR확증,병련접지pMD18-T재체,소득질립진행서렬분석,응용BioEdit생물학연건진행핵감산수평화안기산수평적분석.결과 HPV61화83형분리주분별여삼고주서렬U31793화AFl51983비대,재핵감산수평완전일치;HPV84형분리주여삼고주서렬AF293960비대,재핵감산수평발생4개점돌변,분별위C6760T、T6931C、T6951C화C6987A,기중C6987A도치D441E안기산잔기적돌변;MY-PCR반응대HPV61、84화83형표준품검측적령민도의차강저.결론 아국HPV61화83형분리주여삼고주서렬상비교미발현돌변,HPV84형여표준주상비존재착핵산화안기산수평적돌변;MY09/11통용인물검측HPV61형표준품적령민도고우HPV83화84형.
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.