Typing of human papillomaviruses by consensus polymerase chain reaction and a non-radioactive reverse dot blot hybridization.
A non-radioactive reverse dot blot hybridization method was developed for typing of human papillomavirus (HPV) consensus primer generated polymerase chain reaction (PCR) products in a single test. In the PCR biotin-14-dATP was incorporated into the amplified DNA, which was then used as a probe in hybridization with a membrane, on which different genital HPV types had been immobilized. Of cervical brush samples from women referred to a colposcopy clinic (n = 58) and from women attending a health control program (n = 14) which had been found positive by PCR with consensus HPV primers but negative using primers specific for the HPV types 6, 11, 16, 18, 31, 33 and 35, 25 (43%) and 3 (21%), respectively, could be typed by this method. The additional HPV types found were 34, 39, 40, 45, 52, 53, 56 and 58. Of the samples from the colposcopy clinic (n = 33) and the health control group (n = 11) which could not be typed, 23 and 5, respectively, showed HPV X which cross-hybridized with various HPV types under conditions of low stringency. It is possible to type by this fast and easy method consensus primer-generated PCR products of a wide range of HPV types or to verify the presence of HPV DNA of unknown types.
Forslund
O
O
Department of Medical Microbiology, University of Lund, Malmö General Hospital, Sweden.
Hansson
B G
BG
Bjerre
B
B
D002584:Cervix Uteri / Q000821:virology*; D003001:Cloning, Molecular / Q000379:methods; D016384:Consensus Sequence; D004247:DNA / Q000096:biosynthesis; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis; D005260:Female; D006801:Humans; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012016:Reference Values
D017931:DNA Primers; D004279:DNA, Viral; D004247:DNA