1887

Journal of Medical Microbiology logo

Volume 67, Issue 10

Development and validation of a multiplex reverse transcript real-time PCR for E6/E7 mRNA detection of high-risk human papillomavirus Free

Abstract

Purpose. Human papillomavirus (HPV) E6/E7 mRNA is a more specific marker for cervical lesion screening than HPV DNA. Here, we aimed to develop a new one-step multiplex reverse transcript real-time PCR (MRT-PCR) to detect E6/E7 mRNA from 14 high-risk HPV (hrHPV) genotypes.

Methodology. The analytical sensitivity and specificity of the MRT-PCR system were validated. Its clinical performance was evaluated by comparing the results with bDNA signal amplification assay and histopathological results.

Results. The detection limit of MRT-PCR was 20 to 200 copies per reaction of different HPV genotypes, and no cross-reactivity was observed with any other low-risk HPV or other pathogens commonly found in the female genital tract. Using the bDNA signal amplification assay for comparison, a test on 166 clinical samples showed that the overall agreement between the two methods was 95.18 % and the one-step MRT-PCR was more sensitive. Further, compared with the histopathological results for the 166 clinical samples, the sensitivity and specificity of the MRT-PCR method were 88.9 and 71.6 %, respectively, and the positive rate for hrHPV E6/E7 mRNA increased with the severity of the cervical lesions.

Conclusion. The one-step multiplex RT-PCR for E6/E7 mRNA detection is a simple, fast, universally applicable, sensitive and highly specific method for hrHPV E6/E7 mRNA detection. It is reliable for cervical lesion screening and of potential value in future clinical applications.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): cervical intraepithelial neoplasia , E6/E7 mRNA , human papillomavirus (HPV) and multiplex reverse transcript real-time PCR
© 2018 The Authors
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2018-08-20
2024-10-05
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/content/journal/jmm/10.1099/jmm.0.000824
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Development and validation of a multiplex reverse transcript real-time PCR for E6/E7 mRNA detection of high-risk human papillomavirus
J. Med. Microbiol. 67, 1509 (2018); https://doi.org/10.1099/jmm.0.000824
/content/journal/jmm/10.1099/jmm.0.000824
/content/journal/jmm/10.1099/jmm.0.000824
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