尖锐湿疣患者人乳头瘤病毒6、11型的检测及L1基因的序列分析

Assay and L1 Gene Sequence Analysis of Human Papillomavirus Type 6 and 11 in Condylomata Acuminata

目的 检测尖锐湿疣患者组织中人乳头瘤病毒 (HPV)6型和 11型的感染率，分析 L1晚期基因序列及其蛋白质的氨基酸序列，为基因工程疫苗的研究提供依据。方法 采用 PCR方法测定北京和锦州两地区 34例尖锐湿疣患者病损组织中 HPV6型及 11型的感染率 ,并扩增 L1晚期基因，构建重组测序质粒，双脱氧法测序观察其变异状况。结果 HPV11型感染率为 73.5％ (25/34)， 6型感染率为 44.1％ (15/34),其中两型混合感染率为 17.6％ (6/34)。 11型 L1基因 7～ 8个碱基变异，推导的蛋白质一级结构中可能有 1～ 3个氨基酸变异， 6型 L1基因仅有 3个碱基发生同义突变。结论 检测的 34例患者中以 HPV11型感染为主，兼有 6型和两型的混合感染。 11型 L1基因及蛋白有一定变异， 6型 L1基因仅发生少数碱基同义突变。

To determinate the prevalence of HPV type 6, 11 in condylomata acuminata, and analyze the sequence of L1 gene and its deduced L1 protein . Methods PCR was used to amplify DNA sequences between E6 and E7 of HPV type 6 and 11. The PCR products were identified by the fragment length cleaved with restriction enzyme RsaI. L1 genes were amplified with high fidelity Taq, and cloned DNA sequences were detected by the dideoxynucleotide method. Results In 34 examples of condylomata acuminata, HPV11 was found in 25 examples (73.5％ ), HPV6 in 15(44.1％ ), including HPV6＋ HPV11 in 6(17.6％ ). In comparison with the prototypes there were 7～ 8 point mutations in HPV11 L1 gene and 3 in HPV6L1 gene. L1 encoded protein changes showed up only in 1～ 3 amino acids in the former cases. Conclusion Most of patients tested with condylomata acuminata are mainly infected by HPV11, the rest of them by HPV6 or by co-infection of two mixed types. Sequence analysis of L1 genes indicated that mutations were found in both HPV6 and 11, whereas deduced amino acid mutations in L1 protein were limited in HPV11 only.