title: Development of a fluorescence-based multiplex genotyping method for simultaneous determination of human papillomavirus infections and viral loads.
abstract: BACKGROUND
  Persistent high-risk human papillomavirus (HPV) infection is correlated with an increased risk of developing intraepithelial lesion or malignancy (NILM). The aims of the current study is to establish a method named BioPerfectus Multiplex Real Time (BMRT) HPV assay for simultaneous typing and quantifying HPVs, and to evaluate it by comparison with HPV GenoArray test and PCR-sequencing method, as well as histological status.
  
  
  METHODS
  A total of 817 cervical specimens were evaluated by BMRT method and HPV GenoArray test, using PCR-sequencing method as the reference standard; simultaneously, high-risk HPV-16 and -18 DNA loads were assessed in 443 specimens to investigate the correlation with infection outcomes.
  
  
  RESULTS
  The overall detection coincidence rate between BMRT assay and HPV GenoArray test is 96.6 % and the Kappa value is 0.760. In addition, the sensitivity and positive predictive value of BMRT is 98.4 % and 95.7 % compared with the results detected by PCR-sequencing method, respectively. HPV-16 viral load has a correlation with CINs or worse lesions. By comparing with infected women presenting NILM /cervicitis, the cutoff value for HPV-16 from patients with CINs was 0.827. With this cutoff value, 74.6 % sensitivity and 72.5 % specificity for prediction of HPV-16 infected patients with CINI and higher CIN were achieved. High significance was obtained when comparing the infected women presenting NILM/cervicitis with women either with CIN and cervical carcinomas (p < 0.001).
  
  
  CONCLUSIONS
  The BMRT assay seemed to be a good alternative approach for HR-HPV testing, due to its high level of automation and ability to quantify HPV-16, HPV-18 and other HR-HPVs.
authors.affiliation: Virus Laboratory, The Affiliated Shengjing Hospital, China Medical University, Shenyang, Liaoning, 110004, China. sunzr@sj-hospital.org.
authors.forename: Zhengrong
authors.initials: Z
authors.lastname: Sun
authors.affiliation: Jiangsu Bioperfectus Technologies Limited Company, Jiangsu, 225300, China. zhangrong129@163.com.
authors.forename: Rong
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Jiangsu Bioperfectus Technologies Limited Company, Jiangsu, 225300, China. liuzhonghua@s-sbio.com.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Virus Laboratory, The Affiliated Shengjing Hospital, China Medical University, Shenyang, Liaoning, 110004, China. henryliu17@hotmail.com.
authors.forename: Chao
authors.initials: C
authors.lastname: Liu
authors.affiliation: Jiangsu Bioperfectus Technologies Limited Company, Jiangsu, 225300, China. xiulinli@hotmail.com.
authors.forename: Xiulin
authors.initials: X
authors.lastname: Li
authors.affiliation: Virus Laboratory, The Affiliated Shengjing Hospital, China Medical University, Shenyang, Liaoning, 110004, China. zhgrw@126.com.
authors.forename: Weiqiang
authors.initials: W
authors.lastname: Zhou
authors.affiliation: Virus Laboratory, The Affiliated Shengjing Hospital, China Medical University, Shenyang, Liaoning, 110004, China. 19670445@qq.com.
authors.forename: Lianxia
authors.initials: L
authors.lastname: Yang
authors.affiliation: Virus Laboratory, The Affiliated Shengjing Hospital, China Medical University, Shenyang, Liaoning, 110004, China. ruanq@sj-hospital.org.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Ruan
authors.affiliation: Jiangsu Bioperfectus Technologies Limited Company, Jiangsu, 225300, China. xuzhang@s-sbio.com.
authors.forename: Xu
authors.initials: X
authors.lastname: Zhang
mesh_terms: D052685:Alphapapillomavirus / Q000145:classification* / Q000235:genetics*; D002584:Cervix Uteri / Q000473:pathology / Q000821:virology; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D005838:Genotype; D052162:Human papillomavirus 16 / Q000235:genetics; D052161:Human papillomavirus 18 / Q000235:genetics; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019562:Viral Load; D002578:Uterine Cervical Dysplasia / Q000209:etiology / Q000473:pathology
chemical_list: D004279:DNA, Viral