Α 2-stage, nested-like nucleic acid amplification method (IsoPCR) for the highly sensitive detection of HPV16 and HPV18 DNA. (June 2019)

• Record Type: Journal Article
• Authors: Daskou, M.; Tsakogiannis, D.; Dimitriou, T.G.; Manali, M.; Apti, C.; Amoutzias, G.D.; Mossialos, D.; Kottaridi, C.; Markoulatos, P.
• Is Part Of: Molecular and cellular probes. Volume 45(2019)
• Page Start: 1
• Page End: 7
• Publication Date: 2019-06
• Subjects: IsoPCR -- LAMP -- HPV genotyping -- HPV16 -- HPV18 -- Cervical dysplasia; LAMP Loop-mediated isothermal amplification -- FIP forward inner primer -- BIP backward inner primer -- F3 forward outer primer -- B3 backward outer primer -- LF loop forward primer -- LB loop backward primer -- GuSCN guanidine thiocyanate -- HPV Human papillomavirus.; Molecular probes -- Diagnostic use -- Periodicals; Pathology, Cellular -- Technique -- Periodicals; Cell Biology -- Periodicals; Molecular Biology -- Periodicals; Sondes moléculaires -- Utilisation diagnostique -- Périodiques; Cytopathologie -- Technique -- Périodiques; 572
• DOI: 10.1016/j.mcp.2019.03.003
• Languages: English
• ISSNs: 0890-8508

Abstract

Abstract: Molecular detection of HPV DNA is considered as the gold standard for the diagnosis of cervical disease. Although the molecular assays for the identification of HPV16 and HPV18 have helped identify cervical cancer incidents, they are restricted to specialized laboratories. Thus, we developed a novel 2-stage, nested-like nucleic acid amplification method, named IsoPCR, to amplify the E6 gene of HPV16 and HPV18 with high analytical sensitivity and specificity. The performance of IsoPCR was compared to that of conventional PCR and LAMP. The analytical sensitivity of IsoPCR (1 copy/test) was 10-fold higher than conventional PCR and 25-fold higher than conventional LAMP. IsoPCR displayed significant amplification specificity (100%) and efficiency, as well. In conclusion, IsoPCR is a highly sensitive and specific diagnostic tool and it is suitable for the detection of low copy number of viral DNA in clinical specimens, providing critical information to healthcare providers. Highlights: The IsoPCR assay was developed for the detection of HPV16, HPV18 DNA. The IsoPCR amplify the E6 gene of HPV16 and HPV18 DNA. The assay enables the amplification of 1 copy/reaction of both HPV16, HPV18 DNA. The assay exhibits 100% specificity for the detection of HPV16, HPV18 in clinical samples.