潮州地区人乳头瘤病毒52型E6多态性分析

Analysis of E6 gene polymorphism of type 52 human papillomavirus from Chaozhou area of China

摘要:

目的：明确潮州地区人乳头瘤病毒（Ｈuman papillomavirus，HPV）52型E6编码区的基因多态性。方法：提取101例HPV 52单纯感染者宫颈脱落细胞DNA，通过特异性引物扩增HPV 52-E6编码区全序列，成功获得83份HPV 52-E6 PCR扩增阳性产物，进行测序及BLAST多态性分析。结果：成功获得76条HPV 52-E6基因全序列（447 bp）。该序列与参考株（GQ472848）的一致性为99%。在潮州地区发现11种E6编码区多态性，9种为同义突变，2种有义突变，引起所编码氨基酸改变。根据基因多态性分亚洲群和欧美群2群。亚洲群有7组，第1组，也是最为常见组（50/76）的编码序列与广州（EU924143）和香港（DQ057293）株编码完全匹配，该组称为潮州地方株；第2组（12/76）多态性与广州（EU924144）等报道的E6全序列完全匹配；第3组（仅1例）多态性与浙江大学报道（HM004117）的E6全序列完全匹配；其余4组多态性国际上尚未报道。欧美群有3组，第1组（5/76）多态性与美国（HQ537745）报道的E6全序列完全匹配；第2组（2例）多态性与美国（DQ057291）和德国（X74481）报道的E6全序列完全匹配；第3组（1例）多态性国际上尚未报道。结论：潮州地区HPV 52-E6编码区序列存在多态性，以中国东南部和东南亚国家报道E6编码区序列为主，少数与欧美报道E6全序列完全匹配。为基因疫苗的研制提供了科学依据。

Abstract:

Objective：To study the polymorphism of E6 gene of HPV 52 in Chaozhou. Methods：The DNA was extracted from the cervical exfoliated cells of 101 cases of simple infection of HPV 52. 83 cases of E6 gene complete sequence was amplified success－fully and sequenced. The BLAST 2.0 software online was used for polymorphism study. Results：Totally，76 complete E6 genes were successfully amplified. The similarity was 99% compared with reference strain（GQ472848）. 11 kinds of polymorphism were found，of which 9 kinds were nonsense mutation and the other 2 kinds were sense mutation，which would cause amino acid alteration. All of the 76 cases were divided into Asian cluster and American-European cluster according to the polymorphism of E6 gene. Asian cluster had seven groups. The most common polymorphism was completely matched with the sequence reported in Guangzhou （EU924143） and Hongkong（DQ057293），and this group was named the predominant sequence in Chaozhou area；the variant of the second group was completely matched with the sequence reported in Guangzhou（EU924144）；the variant of the third group was completely matched with the one reported in Zhejiang University reported（HM004117）；while the other 4 groups had never been reported worldwide so far. American-European cluster had three groups. The most common polymorphism was found in 5 different cases，which was com－pletely matched with the sequence reported in America （HQ537745）. The second group polymorphism were found in 2 samples，it was completely matched with sequence reported from America （DQ057291） and Germany （X74481）. The third group was found in one case，which had not been reported before. Conclusion：There is a complicated polymorphism of HPV52 E6 genes in Chaozhou area. The common polymorphism is similar with that reported in the South-east in China or the South-east Asia. The rare sequence matches with American and European reports. This study provides molecular evidence for gene vaccine exploration in Chaozhou.