[Simultaneous detection and typing of human papillomavirus in cervical biopsies using PCR-reverse hybridization].
BACKGROUND
Genotyping of Human papillomavirus (HPV) is an important step in the clinical evaluation of the oncogenic risk associated with HPV infection of cervical mucosa. The purpose of this work was to develop a fast PCR-reverse-hybridization assay (PCR-RH) for the simultaneous detection and genotyping of anogenital HPVs.


METHODS
HPV DNA from cervical biopsies was amplified by consensus primer-PCR. Digoxigenin-labeled PCR products were hybridized to type-specific probes anchored to the surface of plastic microwells and revealed by an ELISA system.


RESULTS
The method was tested on 115 clinical samples (81 koilocytic atypias, 11 CIN1, 10 CIN2, 12 CIN3 and 1 squamous carcinoma). HPV DNA was found in 56.7% koilocytic atypias, in 90.9% of CIN1 and in 100% of CIN2 and higher-grade lesions. Thus, PCR-RH is sensitive, rapid, easy-to-perform and readily applicable to the routine analysis of a large number of samples.
Laconi
S
S
Dipartimento di Scienze Biomediche e Biotecnologie, Università di Cagliari, Cagliari. slaconi@vaxca1.unica.it
Greco
M
M
Milia
G
G
Pellegrini-Bettoli
P
P
Pani
P
P
Laconi
E
E
Rais
M
M
D001706:Biopsy; D002294:Carcinoma, Squamous Cell / Q000473:pathology / Q000821:virology; D002584:Cervix Uteri / Q000473:pathology / Q000821:virology*; D015344:DNA Probes, HPV; D004279:DNA, Viral / Q000032:analysis*; D004076:Digoxigenin; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000473:pathology / Q000821:virology*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D014412:Tumor Virus Infections / Q000175:diagnosis / Q000473:pathology / Q000821:virology*; D002583:Uterine Cervical Neoplasms / Q000473:pathology / Q000821:virology; D002575:Uterine Cervicitis / Q000175:diagnosis / Q000473:pathology / Q000821:virology*; D002578:Uterine Cervical Dysplasia / Q000473:pathology / Q000821:virology
D015344:DNA Probes, HPV; D004279:DNA, Viral; D004076:Digoxigenin