Single-tube Seminested PCR Assay for Detecting Human Papillomavirus in Clinical Samples
Rajan Saini1, *, Jacinta Santhanam2, Nor Hayati Othman3, Deepti Saini1, Thean-Hock Tang4, *
Identifiers and Pagination:Year: 2009
First Page: 106
Last Page: 112
Publisher Id: TOMICROJ-3-106
Article History:Received Date: 19/6/2009
Revision Received Date: 25/6/2009
Acceptance Date: 26/6/2009
Electronic publication date: 22/7/2009
Collection year: 2009
open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.
There is a growing appreciation of the potential value for routine screening for the presence of HPV not only for cervical specimens but also from oral cavity. The purpose of this study was to develop and clinically evaluate a single-tube seminested PCR assay for the detection of HPV. Several parameters such as PCR primers, primer annealing temperature, the number of PCR cycles and concentration of PCR components were optimized. The assay was evaluated using HPV inserts of type 6, 11, 16, 18, 31, 33, 38 and 51. Evaluation of seminested PCR assay was performed with cervical scrapings from 30 patients and buccal swabs from 30 head and neck cancer patients and results were compared with those of two-tube nested PCR. The results were found to be comparable with a total of 60% (36/60) of samples being positive for HPV using the single-tube assay, while 62% (37/60) positivity was found with two-tube PCR assay. We succeeded in developing a single-tube seminested PCR method for HPV DNA detection which is easier than the conventional nested PCR and can be further evaluated as a potential screening tool for detecting HPV in oral and cervical regions.