Single-tube Seminested PCR Assay for Detecting Human Papillomavirus in Clinical Samples
Rajan Saini1, *, Jacinta Santhanam2, Nor Hayati Othman3, Deepti Saini1, Thean-Hock Tang4, *
1 School of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia
2 Department of Biomedical Sciences, Faculty of Allied Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, Kuala Lumpur, 50300, Malaysia
3 Department of Pathology, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia
4 Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 16150 Kubang Kerian, Malaysia, Current Institution: Advanced Medical and Dental Institute (AMDI), Universiti Sains Malaysia, 13200 Bertam, Penang, Malaysia
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.
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* Address correspondence to these authors at the School of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia; Tel: 6097663687; Fax: 6097642026; E-mail: email@example.com and Infectious Disease Cluster, Advanced Medical and Dental, Institute (AMDI), Universiti Sains Malaysia. 13200 Kepala Batas, Penang, Malaysia; Tel: 604-5622888; Fax: 604-5622462; E-mail: firstname.lastname@example.org