Research progress of molecular assay for human papillomavirus
[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 600 000 cases of cervical cancer worldwide,with 340 000 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 capture Ⅱ technology 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.
human papillomaviruscervical cancermolecular assay
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