Rapid detection and typing of human papillomaviruses by consensus polymerase chain reaction and enzyme-linked immunosorbent assay.
A novel method for the detection and typing of human papillomaviruses (HPV) based on consensus polymerase chain reaction (PCR) using MY09/MY11 primers followed by detection of PCR products in a standard microtiter plate format using a recently developed commercially available standardised PCR ELISA kit (Boehringer Mannheim, Germany) was developed. The reliability and feasibility of the method were evaluated on 140 HPV-positive and 85 HPV-negative DNA samples extracted from different archival clinical specimens. Virtually complete agreement between the results of this novel method and the results of previous in-house PCRs and typing method was obtained. The sensitivity level of the novel method, determined by serial log-dilutions of SiHa cells, is about 50 copies of HPV 16. The PCR-ELISA provides the potential for an automated, simple, rapid and accurate test for detection and typing of HPV in diagnostic virological laboratories.
Poljak
M
M
Institute of Microbiology, Medical Faculty of Ljubljana, Slovenia.
Seme
K
K
D001483:Base Sequence; D016384:Consensus Sequence; D004279:DNA, Viral / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005240:Feasibility Studies; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000276:immunology / Q000473:pathology / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured; D014412:Tumor Virus Infections / Q000276:immunology / Q000473:pathology / Q000821:virology*
D004279:DNA, Viral