SIMULTANEOUS DETECTION AND TYPING STRATEGY FOR HUMAN PAPILLOMAVIRUSESBASED ON PCR AND RESTRICTION-ENDONUCLEASE MAPPING

Citation
S. Chen et al., SIMULTANEOUS DETECTION AND TYPING STRATEGY FOR HUMAN PAPILLOMAVIRUSESBASED ON PCR AND RESTRICTION-ENDONUCLEASE MAPPING, BioTechniques, 17(1), 1994, pp. 138
Citations number
11
Categorie Soggetti
Biochemical Research Methods
Journal title
ISSN journal
07366205
Volume
17
Issue
1
Year of publication
1994
Database
ISI
SICI code
0736-6205(1994)17:1<138:SDATSF>2.0.ZU;2-G
Abstract
The aim of this study was to devise a simple and reliable method for s imultaneous detection and typing of genital human papillomaviruses (HP Vs). The method comprises three steps (i) amplification of sample DNA with the L1 consensus primers, (ii) digestion of PCR products with res triction endonuclease RsaI and (iii) hybridization of digested PCR pro ducts with a unique oligonucleotide probe that detects different fragm ents of the six most common genital HPV genotypes, namely, HPV types 6 , 11, 16, 18, 31 and 33. Seventeen clinical specimens were analyzed by this method and compared to Southern blot using full genomic HPV DNA probes and to PCR using type-specific probes. All three tests agreed w ith at least one HPV type; however, the new method identified more add itional HPV types in cases of mixed infections.