J Gynecol Oncol.  2008 Dec;19(4):251-255. 10.3802/jgo.2008.19.4.251.

Comparison between L and E gene amplification analytical methods for human papillomavirus typing

• Cho HB
• Kim YJ
• Kim KT

• KMID: 2173415
• DOI: http://doi.org/10.3802/jgo.2008.19.4.251

Abstract

OBJECTIVE
L and E6/E7 gene amplification analyses were compared to identify human papillomavirus (HPV) infection and verify the HPV type, with the intent to minimize HPV typing errors.
METHODS
L1 gene verified HPV typing was accomplished via polymerase chain reaction (PCR) and membrane assays. Verification of HPV typing via E6/E7 genes was accomplished through nested multiplexed PCR. The results from 104 samples were compared.
RESULTS
The rates of accordance and difference were 35% and 65%, respectively. For 29% of the analyses, nested multiplexed PCR was more diversified than the membrane assay.
CONCLUSION
HPV can be classified into low-risk HPV and high-risk HPV groups. In parallel amplifications of the L and E genes is more efficient for accurate diagnosis in light of the different symptoms and attendant precautions of the risk groups.

Keyword

Human papillomavirus; L1 gene; E6/E7 gene

MeSH Terms

Gene Amplification
Humans
Light
Membranes
Polymerase Chain Reaction

Figure

• 

  Fig. 1 HPV typing by hybridization with L1 gene type-specific probe and nested multiplexed PCR with E6/E7 gene primer sets of cocktails. (A) Membrane assay for HPV typing. GP5+/GP6+ PCR products of 150 bp were selected by electrophoresis on a 2% agarose gel and membrane assay by hybridization with type-specific probe. (B) Nested multiplexed PCR for HPV typing. Primary PCR products were used as the template DNA of each cocktail for nested multiplexed PCR. 1: HPV type 39, 2: HPV type 44, 3: HPV type 58, 4: HPV type 66, 5: HPV type 6, L: 100 bp DNA ladder.

• 

  Fig. 2 Multiple alignment of L gene from types 6, 58, 51, and 66 of human papillomavirus.

Reference

1. Ferlay J, Bray P, Pizani P, Parkin DM. Globocan 2002: Cancer incidence, mortality and prevalence worldwide. 2004. Lyon: IARC Press.

2. Khanna N, Phillips MD. Adherence to care plan in women with abnormal Papanicolaou smears: A review of barriers and interventions. J Am Board Fam Pract. 2001. 14:123–130.

3. Castellsagué X, Muñoz N. Chapter 3: Cofactors in human papillomavirus carcinogenesis - role of parity, oral contraceptives, and tobacco smoking. J Natl Cancer Inst Monogr. 2003. 31:20–28.

4. Bosch FX, Lorincz A, Muñoz N, Meijer CJ, Shah KV. The causal relation between human papillomavirus and cervical cancer. J Clin Pathol. 2002. 55:244–265.

5. Muñoz N, Bosch FX, Castellsagué X, Díaz M, de Sanjose S, Hammouda D, et al. Against which human papillomavirus types shall we vaccinate and screen? The international perspective. Int J Cancer. 2004. 111:278–285.

6. von Krogh G. Management of anogenital warts (condylomata acuminata). Eur J Dermatol. 2001. 11:598–603.

7. Prendiville W, Davies P. Prendiville W, Davis P, editors. The human papillomavirus. The health professionals HPV handbook: Human papillomavirus and cervical cancer. 2004. Oxford: Taylor and Francis;11–26.

8. Luxton J, Shepherd P. Human papillomavirus antigens and T-cell recognition. Curr Opin Infect Dis. 2001. 14:139–143.

9. de Villiers EM, Fauquet C, Broker TR, Bernard HU, zur Hausen H. Classification of papillomaviruses. Virology. 2004. 324:17–27.

10. Smits HL, Tieben LM, Tjong-A-Hung SP, Jebbink MF, Minnaar RP, Jansen CL, et al. Detection and typing of human papillomaviruses present in fixed and stained archival cervical smears by a consensus polymerase chain reaction and direct sequence analysis allow the identification of a broad spectrum of human papillomavirus types. J Gen Virol. 1992. 73:3263–3268.

11. Lungu O, Sun XW, Felix J, Richart RM, Silverstein S, Wright TC Jr. Relationship of human papillomavirus type to grade of cervical intraepithelial neoplasia. JAMA. 1992. 267:2493–2496.

12. Van Den Brule AJ, Walboomers JM, Du Maine M, Kenemans P, Meijer CJ. Difference in prevalence of human papillomavirus genotypes in cytomorphologically normal cervical smears is associated with a history of cervical intraepithelial neoplasia. Int J Cancer. 1991. 48:404–408.

13. Jacobs MV, de Roda Husman AM, van den Brule AJ, Snijders PJ, Meijer CJ, Walboomers JM. Group-specific differentiation between high- and low-risk human papillomavirus genotypes by general primer-mediated PCR and two cocktails of oligonucleotide probes. J Clin Microbiol. 1995. 33:901–905.

14. Sasagawa T, Minemoto Y, Basha W, Yamazaki H, Nakamura M, Yoshimoto H, et al. A new PCR-based assay amplifies the E6-E7 genes of most mucosal human papillomaviruses (HPV). Virus Res. 2000. 67:127–139.

15. Sotlar K, Diemer D, Dethleffs A, Hack Y, Stubner A, Vollmer N, et al. Detection and typing of human papillomavirus by e6 nested multiplex PCR. J Clin Microbiol. 2004. 42:3176–3184.

16. Longworth MS, Laimins LA. Pathogenesis of human papilloviruses in differentiating epithelia. Microbiol Mol Biol Rev. 2004. 68:362–372.

17. Münger K, Howley PM. Human papillomavirus immortalization and transformation functions. Virus Res. 2002. 89:213–228.