Open AccessDNA Microarray Format for Detection and Subtyping of Human Papillomavirus
Author(s) -
Corné H. W. Klaassen,
Clemens Prinsen,
Hanneke A. de Valk,
A. M. Horrevorts,
Marcel Jeunink,
Erik Thunnissen
Publication year - 2004
Publication title -
journal of clinical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.349
H-Index - 255
eISSN - 1070-633X
pISSN - 0095-1137
DOI - 10.1128/jcm.42.5.2152-2160.2004
Subject(s) - amplicon , subtyping , dna microarray , oligonucleotide , digoxigenin , biology , microbiology and biotechnology , hybridization probe , human papillomavirus , dna , papillomaviridae , microarray , polymerase chain reaction , gene , in situ hybridization , gene expression , genetics , medicine , cancer , cervical cancer , computer science , programming language
A new human papillomavirus (HPV) assay using high-density DNA microarrays is described. An HPV DNA fragment from the 3' end of the E1 gene was amplified and digoxigenin labeled by PCR, and the resulting amplicons were hybridized onto type-specific oligonucleotides immobilized on high-density DNA microarrays. For detection, a simple immunohistochemical staining procedure was used with a substrate that has both colorimetric and fluorescent properties. This detection chemistry enables the rapid identification of reactive spots by regular light microscopy and semiquantification by laser scanning. Both single and multiple HPV infections are recognized by this assay, and the corresponding HPV types are easily identified. With this assay, 53 mucosal HPV types were detected and identified. A total of 45 HPV types were identified by a single type-specific probe, whereas the remaining 8 mucosal HPV types could be identified by a specific combination of probes. The simple assay format allows usage of this assay without expensive equipment, making it accessible to all diagnostic laboratories with PCR facilities.