Intended use
Typing of 18 different high risk HPV types. Amplimers originating from the PCR reaction with the primers of this kit or amplimers originating from the EIA kit HPV GP HR can be used for typing.

 

The Genotyping kit HPV GP is an in vitro reverse hybridization strip assay using GP5+/6+ primers for the qualitative identification of the individual high risk types HPV16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73 and 82 in DNA extracted from clinical specimens. The Beta-globin primers are provided to control for sample quality (cervical scrapes, vaginal scrapes, biopsies) and the presence of inhibiting factors. An interpretation for assigning women at risk for developing cervical cancer can not be given on the result of this genotyping assay. The kit is used as an aid in the type specific identification of high risk human papillomavirus infections only.

Summary and explanation
Human papillomaviruses (HPV) are small viruses containing a double-stranded, circular DNA genome of approximately 7,900 base pairs. The viral genome contains early (E) and late (L) genes, as well as an untranslated control region. At present, more than 100 different HPV types have been identified based on differences in DNA sequence. HPV types can be subdivided into mucosal types, which can infect anogenital and oraloropharyngeal mucosa, and cutaneous (skin) HPV types. Among the HPV types infecting the anogenital epithelia, a subset of 18 HPV types have been classified as high-risk or probably high-risk for causing alterations of the cervical mucosa and ultimately cervical cancer in women. The Genotyping kit HPV GP allows an easy and reliable identification of high-risk HPV genotypes using PCR products encompassing a highly conserved L1 sequence that is amplified by the GP5+/6+ PCR primers including a sample quality check by beta globin primers. PCR products can be generated either by the primers that are supplied with the kit or by use of the EIA kit HPV GP HR.

Principle of the procedure
The Genotyping kit HPV GP is based on the reverse hybridization of amplicons obtained by PCR of DNA extracted from clinical specimens. Denatured biotinylated amplicons, resulting from the amplification of a part of the L1 region with GP primers, are hybridized with specific oligonucleotide probes, which are immobilized as parallel lines on membrane strips. After hybridization and stringent washing, streptavidin-conjugated alkaline phosphatase is added and bound to any biotinylated hybrid previously formed. Incubation with BCIP/NBT chromogen yields a purple precipitate and the results can be visually interpreted.

Ordering information
REF: K-72 Genotyping kit HPV GP, version 2 20 tests
(CE registered)

Related products

REF: K-36 EIA kit HPV GP HR 96 tests
(CE registered)

REF: K-90 LMNX kit HPV GP HR 96 tests

References

1) zur Hausen H. Papillomavirus infections-a major cause of human cancers. Biochim. Biophys. Acta 1288, F55-F78 (1996)

2) de Villiers EM, Fauquet C, Broker TR, Berhard HU, zur Hausen H. Classification of papillomaviruses. Virol. 324, 17-27 (2004).

3) Munoz N, Bosch FX, de Sanjose S, Herrero R, Castellsague X, Shah KV et al. Epidemiologic classification of human papillomavirus types associated with cervical cancer. N. Engl. J. Med., 348, 518-527 (2003).

4) van den Brule AJ, Pol R, Fransen-Daalmeijer N, Schouls LM, Meijer CJ, Snijders PJ. GP5+/6+ PCR followed by reverse line blot analysis enables rapid and high-throughput identification of human papillomavirus genotypes. J. Clin. Microbiol. 40, 779-787 (2002).

5) Meijer CJLM, Berkhof J, Castle PE. Hesselink AT, Franco EL, Ronco G, Arbyn M, Bosch FX,Cuzick J, Dillner J,lle D, Heideman AM Snijders PJF Guidelines for human papillomavirus DNA test requirements for primary cervical cancer screening in women 30 years and older. Int. J. Cancer: 124, 516–520 (2009).

6) de Sanjose S, Quint WG, Alemany L, et al. on behalf of the Retrospective International Survey and HPV Time Trends Study Group. Human papillomavirus genotype attribution in invasive cervical cancer: a retrospective cross-sectional worldwide study. Lancet Oncol, 11, 1048-1056 (2010).

7) Carozzi FM, Tornesello ML, Burroni E, Loquercio G, Carillo G, Angeloni C, Scalisi A, Macis R, Chini F, Franco, FM, Rossi PG; Prevalence of Human Papillomavirus Types in High-Grade Cervical Intraepithelial Neoplasia and Cancer in Italy. Cancer Epidemiol Biomarkers Prev; 19(9), 2389–400 (2010).

8) Geraets DT, Heideman DAM, de Koning MNC, Snijders PJF, Meijer CJLM, van Doorn LJ, Quint WGV. High genotyping concordance between the digene HPV Genotyping RH Test and the Reverse Line Blot genotyping assay on GP5+/6+-PCR products. J. Clin. Virol, 46(S3), S16–20 (2009).