Development and validation of a multiplex reverse transcript real-time PCR for E6/E7 mRNA detection of high-risk human papillomavirus
Autor: | Ren Xuyi, Pan Caixia, Jing Zhou, Jiangfeng Lyu |
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Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Microbiology (medical) Papillomavirus E7 Proteins Biology Real-Time Polymerase Chain Reaction Cervical intraepithelial neoplasia Sensitivity and Specificity Microbiology 03 medical and health sciences 0302 clinical medicine Genotype medicine Humans Multiplex RNA Messenger Human papillomavirus Papillomaviridae Messenger RNA Papillomavirus Infections Clinical performance Oncogene Proteins Viral General Medicine medicine.disease Molecular biology Hpv testing 030104 developmental biology Real-time polymerase chain reaction 030220 oncology & carcinogenesis RNA Viral Female Multiplex Polymerase Chain Reaction |
Zdroj: | Journal of Medical Microbiology. 67:1509-1514 |
ISSN: | 1473-5644 0022-2615 |
Popis: | 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. |
Databáze: | OpenAIRE |
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