论文部分内容阅读
目的研究广西地区女性人乳头瘤病毒感染情况及HPV16型致癌蛋白变异情况。方法收集广西地区2012年1月-2015年12月8 062例女性就诊患者宫颈拭子标本,采用Premier5.0软件依据Genbank基因库设计引物,PCR扩增HPV基因,反应完成后放置水浴,进行HPV导流杂交,肉眼观察见蓝紫色圆点为阳性,并根据HPV分型图进行分型。PCR产物纯化后双向测序,与标准株进行比对。结果 8 062例患者中有1 524例感染HPV,共计分离出的2058株HPV病毒。其中分离出高危型病毒1 667株,占81.00%;低危型391株,占19.00%。1524例HPV感染者中单一HPV感染1 053例,占69.09%;双重感染411例,占26.97%;三重感染57例,占3.74%;四重感染3例,占0.20%。将患者按年龄分为≤30岁、31-40岁、41-50岁和≥51岁4个年龄组,感染例数和感染率分别为412例和16.27%,656例和22.19%,342例和16.09%,114例和25.39%。569例患者的宫颈拭子标本扩增出HPV致癌蛋白E6基因,其中351株发生基因位变,占61.69%,涉及到15个碱基位16种碱基发生突变,导致11种氨基酸突变;569株扩增出HPV致癌蛋白E7基因,其中307株发生基因位变,占53.95%,涉及到14个碱基位14种碱基发生突变,导致9种氨基酸突变。讨论不同HPV亚型编码外壳蛋白基因有较大区别,具有科目特异性,且在不同亚型间无相互交叉保护性抗体。因而不同HPV亚型可诱发多重感染,并导致疾病发生。
Objective To study the prevalence of human papillomavirus (HPV) infection and the variation of HPV16 oncoprotein in Guangxi area. Methods Cervical swab specimens from 862 female patients were collected from January 2012 to December 2015 in Guangxi. PCR was used to amplify the HPV gene by PCR using Premier5.0 software. After the reaction was completed, the cells were placed in a water bath for HPV Diversion hybridization, macroscopic observation of the blue-violet dots are positive, and according to the HPV subtype. PCR products were purified after two-way sequencing, compared with the standard strains. Results A total of 1 524 HPV infections were detected in 8 062 patients, totally 2058 strains of HPV were isolated. Among them, 1 667 strains of high-risk virus were isolated, accounting for 81.00%; 391 strains were low-risk, accounting for 19.00%. Among 1524 cases of HPV infection, 1 053 cases were single HPV infection, accounting for 69.09%; 411 cases were double infection (26.97%), 57 cases were triple infection (3.74%) and 3 cases were tetrad infection (0.20%). The patients were divided into four age groups of ≤30 years old, 31-40 years old, 41-50 years old and ≥51 years old by age. The number of cases and the infection rate were 412 cases and 16.27%, 656 cases and 22.19%, respectively, and 342 cases And 16.09%, 114 and 25.39% respectively. 569 cases of patients with cervical swab specimens amplified HPV oncoprotein E6 gene, of which 351 strains occurred genetic loci, accounting for 61.69%, involving 15 base 16 mutations occurred, resulting in 11 kinds of amino acid mutations; 569 Strain amplified HPV E7 oncogene protein, of which 307 strains occurred genetic change, accounting for 53.95%, involving 14 base 14 mutations occurred, resulting in 9 kinds of amino acid mutations. Discuss different HPV subtypes coding coat protein gene has a larger difference, with the subject-specific, and no cross-protection antibodies in different subtypes. Thus different HPV subtypes can induce multiple infections and lead to disease.