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[目的]建立灵敏而稳定的人乳头状瘤病毒(human papillomavirus,HPV)58型病毒含量与整合状态检测方法,并探讨HPV58型在我国粤东地区妇女宫颈癌中的感染特征。[方法]PCR扩增HPV58E2和E6基因,PCR产物纯化,连接入T载体,构建标准品;检测30例宫颈癌和40例对照组织样本中HPV58的病毒含量和整合状态。[结果]成功构建含有HPV58E2或E6片段的T载体质粒作为检测HPV58型的病毒含量和判断感染状态标准品。HPV58型在宫颈癌中的感染率和病毒含量较正常宫颈组织均高,并且HPV58在粤东地区宫颈癌中的感染状态以整合型(66.67%)和混合型(33.33%)为主,和正常宫颈组织中感染状态分布有显著差异。[结论]采用荧光定量PCR法所建立的HPV58型病毒含量和整合状态的检测方法具有灵敏性和稳定性,HPV58型在粤东地区宫颈癌中的感染以整合状态为主。
[Objective] To establish a sensitive and stable method to detect the content and integrated status of human papillomavirus (HPV) virus type 58 and investigate the infection characteristics of HPV58 in women with cervical cancer in the eastern part of China. [Methods] The HPV58E2 and E6 genes were amplified by PCR. The PCR products were purified and ligated into T vector to construct standard samples. The virus content and integration status of HPV58 in 30 cases of cervical cancer and 40 cases of control tissues were detected. [Results] The T vector plasmid containing HPV58E2 or E6 fragment was successfully constructed as a standard for detecting the virus content of HPV58 and determining the infection status. The infection rate and virus content of HPV58 in cervical cancer were higher than those in normal cervical tissue. HPV58 infection in cervical cancer in eastern Guangdong Province was mainly integrated (66.67%) and mixed (33.33%), and normal Cervical tissue infection in the distribution of significant differences. [Conclusion] The detection method of HPV58 virus content and integrated status established by fluorescence quantitative PCR method is sensitive and stable. The infection of HPV58 in cervical cancer in the eastern part of Guangdong Province is characterized by the integrated status.