潮州地区人乳头瘤病毒52型L1基因多态性分析

Analysis of L1 gene polymorphism of human papillomavirus type 52 from Chaozhou area of China

目的研究潮州地区优势型别人乳头瘤病毒(HPV)52型L1基因多态性。为基于L1基因工程疫苗的设计和应用提供理论依据。方法采用核酸分子快速导流杂交分型技术(HybriMax)检出HPV52,提取101例HPV52单纯感染者宫颈脱落细胞DNA,通过4对特异性引物分别扩增HPV52-L1编码区全序列,进行测序及BLAST多态性分析。结果本研究成功获得73条HPV52L1基因全序列(1590bp)。该序列与天津参考株(GQ472848)的一致性为99%。在潮州地区发现的所有L1基因序列中有31个碱基是可发生变异的,其中25种为同义突变,6种为有义突变。根据基因多态性分东亚群和欧洲群2群。东亚群有13组,其中CZ52A105是本地最常见的变异株,占61.64%(45/73),该组称为潮州地方株,其编码序列与泰国清迈报道的IN141070株(HQ537743)完全匹配,其余各组(21/73)多态性国际上均尚未报道,他们均与天津报道的TJ49-52(GQ472848)株以及泰国报道的IN141070(HQ537743)株亲缘关系最近,有1～6个碱基不同。欧洲群有2组为CZ52E928(2/73)和CZ52E136(5/73),其基因序列分别与德国REF(X74481)株和北美洲Qv03719(HQ537745)株的L1全序列完全匹配。结论潮州地区HPV52-L1基因多态性与中国东南部和东南亚国家报道的类似,CZ52A105是潮州地方株。

Objective Purpose To study the polymorphism of L1 gene of human papillomavirus（HPV）subtype 52 in Chaozhou area,and provide the useful data for designing and applying of L1 gene relied vaccine. Methods HPV-52 was detected hy HybriMax. The DNA was extracted from the cervical exfoliated cells of 101 cases of simple infection of HPV 52. The complete L1 gene sequence was amplified with four pairs of primers,and then sequenced in Invitrogen Bi- otechnology Co. Ltd. The BLAST 2.0 software online were used to analyze and identify mismatches. Results Totally, 73 complete L1 genes were successfully amplified and sequenced. The similarity was 99% compared with reference strain （GQ472848）. 31 nucleotide acids were variable,25 of them were nonsense mutation,the other 6 were sense mutation. All of 73 cases were divided into East Asian and Europe lineage according to the polymorphism of L1 gene. East Asian cluster include 13 variants, CZ52A105 represent the most common variant（61. 64%, 45/73）, this variant was named ＂Chaozhou isolate＂, its sequence was completely matched with isolate IN141070（HQ537743） reported in Thailand, while the other 12 variants have never been reported worldwide so far, their sequence was the closest homology with sequence reported in Tianjin TJ49-52（GQ472848）and Thailand IN141070（HQ537743）,one to.six mismatches was found. Europe cluster included two kinds of variants, which were isolate CZ52E928 （2/73）and CZ52E136 （5/73）, their sequence were completely matched with sequence reported in German REF （X74481） and Guanacaste Qv03719 （HQ537745）respec- tively. Conclusion The most common polymorphism of HPV52 L1 genes was similar with that reported in the south- east China or other south-east Asia countries,CZ52A105 was named the predominant sequence in Chaozhou area. This study provided molecular evidence for gene vaccine exploration in Chaozhou.