Document Type : Original Article
Authors
1 Department of biology, Faculty of science, Arak university, Arak, Iran
2 Department of Cellular and Molecular Tehran Medical Sciences Branch,lslamic Azad University
Abstract
The causalGiven the known relationship between the HPV infection and some malignancies, it is critical to develop methods for quick detection and quantitation of certain HPV types while encountering a suspected lesion. Early HPV detection is greatly important in monitoring and treating the disease development and progression. Detection of the viral DNA using PCR is the standard, noninvasive method for detecting cervical HPV infection. In the present study, we intended to develop a TaqMan genotyping assay that targets two types of high-risk HPV types (HPV 16 & 18) and two of the low-risk types (6 & 11).
The study included 75 samples positive for HPV, of which 37 were positive for HPV types 16 and 18, while 38 were positive for HPV types 6 and 11. The samples had been confirmed by a reference kit before. The samples underwent real-time PCR. Each reaction consisted of the 1X CAPITAL™ qPCR Probe Master Mix, specific primer pairs for HPV, and fluorescent-tagged probes.
According to our findings, all the samples genotyped using this method were compatible with the results by the reference kit, which was remarkable.
In conclusion, our type-specific approach based on real-time PCR could detect the entire samples positive for four types of HPV.
Keywords
H: Classification of papillomaviruses. Virology 2004, 324:17-27.
2. zur Hausen H: Papillomaviruses and cancer: from basic studies
to clinical application. Nat Rev Cancer 2002, 2:342-50.
3. Palefsky J: Human papillomavirus and anal neoplasia. Curr
HIV/AIDS Rep 2008, 5:78-85.
4. Dianzani C, Calvieri S, Pierangeli A, Degener AM: Identification
of human papilloma viruses in male dysplastic genital lesions.
New Microbiol 2004, 27:65-9.
5. Herrero R, Castellsague X, Pawlita M, Lissowska J, Kee F,
Balaram P, Rajkumar T, Sridhar H, Rose B, Pintos J, et al: Human
papillomavirus and oral cancer: the International Agency for
Research on Cancer multicenter study. J Natl Cancer Inst 2003,
95:1772-83.
6. Schwartz SM, Daling JR, Doody DR, Wipf GC, Carter JJ,
Madeleine MM, Mao EJ, Fitzgibbons ED, Huang S, Beckmann
AM, et al: Oral cancer risk in relation to sexual history and
evidence of human papillomavirus infection. J Natl Cancer Inst
1998, 90:1626-36.
7. D’Souza G, Kreimer AR, Viscidi R, Pawlita M, Fakhry C, Koch
WM, Westra WH,
Gillison ML: Case-control study of human papillomavirus and
oropharyngeal cancer. N Engl J Med 2007, 356:1944-56.
8. Munoz N, Bosch FX, de Sanjose S, Herrero R, Castellsague
X, Shah KV, Snijders PJ, Meijer CJ: Epidemiologic classification
of human papillomavirus types associated with cervical cancer. N
Engl J Med 2003, 348:518-27.
9. Swan DC, Tucker RA, Tortolero-Luna G, Mitchell MF,
Wideroff L, Unger ER, Nisenbaum RA, Reeves WC, Icenogle JP:
Human papillomavirus (HPV) DNA copy number is dependent on
grade of cervical disease and HPV type. J Clin Microbiol 1999,
37:1030-4.
10. Kreimer AR, Clifford GM, Snijders PJ, Castellsague X,
Meijer CJ, Pawlita M, Viscidi R, Herrero R, Franceschi S: HPV16
semiquantitative viral load and serologic biomarkers in oral and
oropharyngeal squamous cell carcinomas. Int J Cancer 2005,
115:329-32.
11. Tucker RA, Unger ER, Holloway BP, Swan DC: Real-
time PCR-based fluorescent assay for quantitation of human
papillomavirus types 6, 11, 16, and 18. Mol Diagn 2001, 6:39-47.
12. Swan DC, Tucker RA, Holloway BP, Icenogle JP: A sensitive,
type-specific, fluorogenic probe assay for detection of human
papillomavirus DNA. J Clin Microbiol 1997, 35:886-91.
13. Peitsaro P, Johansson B, Syrjanen S: Integrated human
papillomavirus type 16 is frequently found in cervical cancer
precursors as demonstrated by a novel quantitative real-time PCR
technique. J Clin Microbiol 2002, 40:886-91.
14. Nagao S, Yoshinouchi M, Miyagi Y, Hongo A, Kodama J,
Itoh S, Kudo T: Rapid and sensitive detection of physical status
of human papillomavirus type 16 DNA by quantitative real-time
PCR. J Clin Microbiol 2002, 40:863-7.
15. Josefsson A, Livak K, Gyllensten U: Detection and quantitation
of human papillomavirus by using the fluorescent 5’ exonuclease
assay. J Clin Microbiol 1999, 37:490-6.
16. Gravitt PE, Peyton C, Wheeler C, Apple R, Higuchi R,
Shah KV: Reproducibility of HPV 16 and HPV 18 viral load
quantitation using TaqMan real-time PCR assays. J Virol Methods
2003, 112:23-33.
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