人乳头瘤病毒分子检测方法研究进展

Research progress of molecular assay for human papillomavirus

[背景]人乳头瘤病毒(human papillomavirus,HPV)是最常见的性传播病原体之一.高危型HPV的持续感染与高度宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)及宫颈癌的发生密切相关,70%的宫颈癌病例是由HPV-16和HPV-18两种高危型引起的.HPV分子检测具备灵敏度高、检测时间短和临床适用性好的技术优势,目前已成为宫颈癌筛查的主要方案.[进展]本文从检测技术原理、检测技术的性能评价、检测技术的优势与不足等方面对HPV分子检测技术研究进展进行综述,为临床宫颈癌筛查工作选择合适的HPV检测及基因分型技术提供支持.[展望]选择适合的HPV检测技术用于宫颈癌筛查需要全面考虑每种技术的应用场景、检测能力和成本负担.将宫颈癌筛查与HPV疫苗接种和宫颈治疗相结合,有望降低宫颈癌的发病率和死亡率,并实现世界卫生组织在全球范围内加速消除宫颈癌的最终目标.

[Objective]Human papillomavirus(HPV)is one of the most common sexually transmitted pathogens and a major cause of cervical cancer.Global cancer research data showed that HPV persistent infection has caused more than 600000 cases of cervical cancer worldwide,with 340000 fatalities in 2020.On November 17,2020,the World Health Organization(WHO)launched a global strategy to accelerate the elimination of cervical cancer,with three actions of HPV vaccination,cervical cancer screening,and cervical disease treatment.Cervical cancer screening is an important measure in this strategy,with cytological testing,HPV testing,and colposcopy tissue biopsy being the most common technologies for cervical cancer diagnosis.Compared to cytological testing and colposcopy tissue biopsy,HPV testing has advantages of good detection sensitivity,high detection throughput,and convenient operation.HPV testing also shows great potentiality for supporting detection model using self-sampling which can improve the coverage of cervical cancer screening population,especially in remote areas.For these advantages,the WHO currently recommends HPV testing as the primary tool for cervical cancer screening.[Progress]Molecular assay for HPV can be performed using many technologies,categorized into two major platforms:nucleic acid hybridization platform and nucleic acid amplification platform.Beyond these two platforms,new technologies have also been applied in molecular assay for HPV,such as next generation sequencing(NGS)technology and in situ hybridization of mRNA technology.The detection targets for HPV molecular assay mainly includes E6 and E7 genes related to the development of cervical cancer,or the L1 gene coding the capsid protein of HPV.Other genes have also been reported as detection targets in HPV molecular assay.Then nucleic acid hybridization platform includes three typical technologies.The first is the hybrid captureⅡtechnology that has the advantage of low instrument requirement and no risk of PCR contamination.The second is the invader technology noted for its good specificity and ability to reflect the transcription level of HPV E6 and E7 genes.The third is the branched DNA technology that eliminates the need for HPV RNA purification and reverse transcription PCR,which reduces the pre-treatment time in the assay.However,nucleic acid hybridization platform requires extensive hand-on operations and is unable to provide information on HPV genotypes.The nucleic acid amplification platform includes the following six typical technologies:real-time PCR technology(known for its good sensitivity,short detection time,and high detection throughput),reverse dot hybridization technology(cost-effective and proficient in enabling HPV full genotyping),fluorescence signal interpretation of gene chip technology(exhibiting stable fluorescence signals and high throughput for HPV genotyping),flow cytometry fluorescence hybridization technology(enhancing the hybridization efficiency of fluorescence probes and targets with similar advantages for high throughput of HPV genotyping),transcription-mediated amplification technology(noted for its good specificity and ability to reflect the transcription level of HPV E6 and E7 genes),and melting curve analysis technology(increasing the detection throughput for HPV genotyping in single fluorescent PCR reaction and offering parallel advantages to real-time PCR technology.NGS technology and in situ hybridization of mRNA technology have also been applied in HPV molecular assay.NGS performs large-scale parallel sequencing,providing comprehensive information including HPV sequences and relative abundance of all HPV genotypes in co-infection HPV sample.In situ hybridization of mRNA technology can directly observe the distribution of HPV mRNA in cervical epithelial cells,and is expected to be a high specificity assay for cervical cancer screening.[Perspective]HPV infection is an important risk factor leading to a variety of human epithelial cytopathies,and molecular assay for HPV has great significance for cervical cancer screening.The selection of suitable HPV detection technology for cervical cancer screening requires comprehensive consideration of the application scenarios,detection capability,and cost burden of each technology.Cervical cancer screening combined with HPV vaccination and cervical treatment holds promise for reducing the morbidity and mortality of cervical cancer and achieving the ultimate goal of the global strategy to accelerate cervical cancer elimination.