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  • Title: Development and validation of a multiplex reverse transcript real-time PCR for E6/E7 mRNA detection of high-risk human papillomavirus.
    Author: Pan C, Zhou J, Lyu J, Ren X.
    Journal: J Med Microbiol; 2018 Oct; 67(10):1509-1514. PubMed ID: 30124401.
    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.
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