Yang Li-Juan, Yue Yao-Fei, Chen Jun-Ying, Pan Yue, Zhao Yu-Jiao, Ma Shao-Hui, Sun Qiang-Ming
Chinese Academy of Medical Sciences and Peking Union Medical College, Institute of Medical Biology, Kunming 650118, China.
Bing Du Xue Bao. 2012 Nov;28(6):645-51.
To investigate E6 and E7 gene variations of human papillomavirus type 16 in Yunnan Province, DNA was extracted from 2000 gynecological outpatient samples. For Human papillomavirus (HPV) genotyping, the genomic DNA was first amplified by the consensus MY09/MY11 primer pair followed by nested PCR with GP5+/GP6+ primers, then the PCR products were subjected to direct DNA sequencing. A total of 20 HPV-16 viral DNAs were identified. E6 and E7 genes of HPV-16 viral DNA were then amplified using E6 and E7 specific primers, the PCR products were purified and sequenced. The results showed that mutations were found at nucleotide position 178 of HPV-16 E6 gene in 10 cases,the mutation rate was 50%; For HPV-16 E7 gene, the mutations were found at nucleotide position 647 in 10 cases; the mutation rate was 50%. Phylogenetic analysis showed that Asian (As) variants of HPV-16 were predominated in Yunnan, China. None of African-1, African-2 variants of HPV-16 was found in this region.
为研究云南省人乳头瘤病毒16型(HPV-16)的E6和E7基因变异情况,从2000份妇科门诊样本中提取DNA。对于人乳头瘤病毒(HPV)基因分型,首先用通用引物MY09/MY11对基因组DNA进行扩增,然后用GP5+/GP6+引物进行巢式PCR,之后对PCR产物进行直接DNA测序。共鉴定出20份HPV-16病毒DNA。然后用E6和E7特异性引物扩增HPV-16病毒DNA的E6和E7基因,对PCR产物进行纯化和测序。结果显示,10例样本中HPV-16 E6基因第178位核苷酸发生突变,突变率为50%;对于HPV-16 E7基因,10例样本中第647位核苷酸发生突变,突变率为50%。系统发育分析表明,HPV-16亚洲(As)变异株在中国云南占主导地位。该地区未发现HPV-16的非洲-1、非洲-2变异株。