Single-tube real-time nested polymerase chain reaction for detecting human papillomavirus DNA

Strauss, Susanne, Jordens, J. Zoe, Desselberger, Ulrich and Gray, Jim J. (2000) Single-tube real-time nested polymerase chain reaction for detecting human papillomavirus DNA. Diagnostic Molecular Pathology, 9 (3). pp. 151-157. ISSN 1052-9551

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Abstract

A single-tube real-time nested polymerase chain reaction (PCR) was developed to detect human Papillomavirus (HPV) DNA in a closed tube system. The oligonucleotide primers MY09/MY11 and GP5+/GP6+ were included in contiguous reactions, thus eliminating the need to transfer first round PCR product into a second tube. The sensitivity and specificity of the optimized single-tube nested PCR were comparable with that achieved by two separate reactions on a conventional thermal block system using serial dilutions derived from plasmids containing DNA of 20 HPV types. A minimum of 10 copies of HPV types 11 and 16 DNA could be detected by both systems. In clinical samples, HPV types 1A, 2, 3, 5, 6-8, 10, 11, 14, 16, 17, 18, 20, 31, 33, 35, 39, 45, 49, 50, 52-54, 57, 62, 66, 70, CP8304 and LVX82/MM7 could be detected by both PCR methods. A total of 145 samples collected from patients were tested for the presence of HPV DNA with the two PCR systems; 124 (86.1%) of 144 samples gave concordant results in both assays. The HPV DNA positive PCR amplicons were typed and concordant results were obtained in 47 of 67 positive samples tested in both amplicons. In samples containing multiple HPV types at least one type was common to both amplicons.

Item Type: Article
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
Related URLs:
Depositing User: Rhiannon Harvey
Date Deposited: 19 May 2011 12:42
Last Modified: 21 Apr 2020 21:12
URI: https://ueaeprints.uea.ac.uk/id/eprint/31065
DOI:

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