Development and validation of a multiplex qPCR assay for detection and relative quantification of HPV16 and HPV18 E6 and E7 oncogenes

被引：0
作者：
Alexia Bordigoni
Anne Motte
Hervé Tissot-Dupont
Philippe Colson
Christelle Desnues
机构：
[1] UMR Microbes,Aix
[2] Evolution,Marseille Université, IRD 198, Assistance
[3] Phylogeny and Infections (MEPHI),Publique des Hôpitaux de Marseille
[4] Mediterranean Institute of Oceanography (MIO),Aix
[5] IHU Méditerranée Infection,Marseille Université, Université de Toulon, CNRS, IRD
来源：
Scientific Reports | / 11卷
关键词：
D O I：
暂无
中图分类号：
学科分类号：
摘要：
Human papillomaviruses (HPV) play a key role in promoting human anogenital cancers. Current high-risk HPV screening or diagnosis tests involve cytological or molecular techniques mostly based on qualitative HPV DNA detection. Here, we describe the development of a rapid quantitative polymerase chain reaction (qPCR) detection test of HPV16 and HPV18 oncogenes (E6 and E7) normalized on human gene encoding GAPDH. Optimized qPCR parameters were defined, and analytical specificities were validated. The limit of detection was 101 for all genes tested. Assay performances were evaluated on clinical samples (n = 96). Concordance between the Xpert HPV assay and the triplex assay developed here was 93.44% for HPV16 and 73.58% for HPV18. HPV co-infections were detected in 15 samples. The systems developed in the present study can be used in complement to traditional HPV tests for specifically validating the presence of HPV16 and/or HPV18. It can also be used for the follow-up of patients with confirmed infection and at risk of developing lesions, through the quantification of E6 and E7 oncogene expression (mRNA) normalized on the GAPDH expression levels.
引用
收藏
相关论文
共 86 条
  • [1] Doorbar J(2015)Human papillomavirus molecular biology and disease association Rev. Med. Virol. 25 2-23
  • [2] Egawa N(2009)Human papillomaviruses: Genetic basis of carcinogenicity Public Health Genom. 12 281-290
  • [3] Griffin H(2004)Classification of papillomaviruses Virology 324 17-27
  • [4] Kranjec C(2016)Human papillomavirus infection Dis. Mon. 65 661-666
  • [5] Murakami I(2016)Human papillomavirus-associated cancers—United States, 2008–2012 MMWR Morb. Mortal. Wkly. Rep. 141 664-670
  • [6] Burk RD(2017)Worldwide burden of cancer attributable to HPV by site, country and HPV type Int. J. Cancer 168 123-137
  • [7] Chen Z(2008)Persistent human papillomavirus infection and cervical neoplasia: A systematic review and meta-analysis Am. J. Epidemiol. 8 405-415
  • [8] Doorslaer KV(2008)Human papillomavirus E6/E7 mRNA testing as a predictive marker for cervical carcinoma Expert Rev. Mol. Diagn. 61 134-144
  • [9] de Villiers E-M(2018)Understanding the HPV integration and its progression to cervical cancer Infect. Genet. Evolut. 7 11-22
  • [10] Fauquet C(2007)The natural history of cervical HPV infection: Unresolved issues Nat. Rev. Cancer 248 76-79
123456789