Intended use
Typing of 24 low-, intermediate- and high risk HPV types.

The RHA kit HPV SPF10-LiPA25, version 1 is an in vitro reverse hybridization strip assay for the qualitative identification of DNA from Human Papillomavirus (HPV) genotypes 6, 11, 16, 18, 31, 33-35, 39, 40, 42-45, 51-54, 56, 58, 59, 66, 68, 70, and 74. Based on the high analytical sensitivity of the assay, this kit is particularly suited for epidemiological and vaccinerelated studies, using both cervical scrapes and biopsy specimens. This kit is validated to be used in combination with the DNA ELISA kit HPV SPF10, version 1.

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 [1]. At present, more than 100 different HPV types have been identified based on differences in DNA sequence [2]. 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 [2, 3] for causing alterations of the cervical mucosa and ultimately cervical cancer in women. The RHA kit SPF10-LiPA25, version 1 allows an easy and reliable identification of HPV genotypes using SPF10 PCR products encompassing a highly conserved L1 sequence.

Principles of the procedure
The RHA kit HPV SPF10-LiPA25, version 1 is based on the reverse hybridization principle. Denatured biotinylated amplicons, resulting from the amplification of a part of the L1 region with SPF10 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
The kit is available for research- and epidemiological studies
REF: S-1071 RHA Kit HPV SPF10-LiPA25, version 1 50 tests
REF: S-1026 RHA Kit HPV SPF10-LiPA25, version 1 500 tests

Related products
REF: K-27 DNA ELISA KIT HPV SPF10, version 1 96 tests
REF: K-27-4 DNA ELISA KIT HPV SPF10, version 1 384 tests

1) Kleter et al. (1998), Novel short-fragment PCR assay for highly sensitive broad-spectrum detection of anogenital human papillomaviruses. Am J Pathol 153, 1731-9.

2) Kleter et al. (1999), Development and clinical evaluation of a highly sensitive PCR-reverse hybridization line probe assay for detection and identification of anogenital human papillomavirus. J Clin Microbiol 37, 2508-17.

3) Quint et al. (2001), Comparative analysis of human papillomavirus infections in cervical scrapes and biopsy specimens by general SPF(10) PCR and HPV genotyping. J Pathol 194, 51-8.

4) van Doorn et al. (2002), Genotyping of human papillomavirus in liquid cytology cervical specimens by the PGMY line blot assay and the SPF(10) line probe assay. J Clin Microbiol 40, 979-83.

5) van der Graaf et al. (2002), Human papillomavirus and the long-term risk of cervical neoplasia. Am J Epidemiol 156, 158-64.

6) Shin et al. (2004), Prevalence and determinants of genital infection with papillomavirus, in female and male university students in Busan, South Korea. J Infect Dis 190, 468-76.

7) Harper et al. (2004), Efficacy of a bivalent L1 virus-like particle vaccine in prevention of infection with human papillomavirus types 16 and 18 in young women: a randomised controlled trial. Lancet 2004 364, 1757-65.

8) van Ham et al. (2005), comparison of two commercial assays for detection of human papillomavirus (HPV) in cervical scrape specimens: validation of the Roche AMPLICOR HPV test as a means to screen for HPV genotypes associated with a higher risk of cervical disorders. J Clin Microbiol 43, 2662-7.

9) Harper et al. (2006), Sustained efficacy up to 4-5 years of a bivalent L1 virus-like particle vaccine against human papillomavirus types 16 and 18: follow-up from a randomised control trial. Lancet 367, 1247-55.

10) Safaeian et al. (2007), Comparison of the SPF10-LiPA system to the Hybrid Capture 2 Assay for detection of carcinogenic human papillomavirus genotypes among 5,683 young women in Guanacaste, Costa Rica. J Clin Microbiol 45, 1447-54.

11) de Koning et al. (2008), Prevalence of mucosal and cutaneous human papillomaviruses in different histologic subtypes of vulvar carcinoma. Mod Pathol 21, 334-44.

12) Quint et al. (2009), Comprehensive analysis of Human Papillomavirus and Chlamydia trachomatis in in-situ and invasive cervical adenocarcinoma. Gynecol Oncol 114, 390-4.

13) Lenselink et al. (2009), Detection and genotyping of human papillomavirus in self-obtained cervicovaginal samples by using the FTA cartridge: new possibilities for cervical cancer screening. J Clin Microbiol 47, 2564-70.

14) Sanjose et al. (2010), Human papillomavirus genotype attribution in invasive cervical cancer: a retrospective cross-sectional worldwide study. Lancet Oncol 11, 1048-56.

15) Quint et al. (2010), HPV genotyping and HPV16 variant analysis in glandular and squamous neoplastic lesions of the uterine cervix. Gynecol Oncol 117, 297-301.

16) Quint et al. (2012), One virus, one lesion, individual components of CIN lesions contain a specific HPV type. J Pathol 227, 62-71.