Detection of the human papilloma virus type 16 mRNA-transcripts in cytological abnormal scrapings.
The RNA-polymerase chain reaction (PCR) was carried out on cervical scrapings to detect and analyze transcripts from the E6-E7 open reading frames (ORF) of human papilloma virus type 16 (HPV16). The method, described previously for cervical squamous carcinomas and cervical intraepithelial neoplasias, was adapted to cervical scrapings. A primer set and two different probes specific for the E6-E7 ORFs were selected. One of the probes was able to detect the amplification products from the full length, the major, and the minor transcripts whereas the other was specific for the major transcript only. To check the quality of the mRNA in the cervical scrapings, a primer set and a probe specific for the human keratin mRNA were selected. A group of 17 abnormal cytological cervical scrapes, which were positive for HPV16 DNA, was analyzed. In this group the human papilloma virus was not always transcriptionally active, as HPV16 mRNA transcripts were detected only in about one-half (8/17) of the samples. These findings suggest that the RNA-PCR method on cervical scrapings may be very useful for epidemiological studies on the role of transcriptionally active/inactive HPV16 genes in the pathogenesis of an HPV16 infected lesion.
Falcinelli
C
C
Department of Research and Development, Diagnostisch Centrum SSDZ, Delft, The Netherlands.
Claas
E
E
Kleter
B
B
Quint
W G
WG
D001483:Base Sequence; D002584:Cervix Uteri / Q000382:microbiology*; D015344:DNA Probes, HPV; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D014158:Transcription, Genetic; D014412:Tumor Virus Infections / Q000150:complications / Q000382:microbiology; D002583:Uterine Cervical Neoplasms / Q000209:etiology; D002575:Uterine Cervicitis / Q000150:complications / Q000382:microbiology; D014626:Vaginal Smears
D015344:DNA Probes, HPV; D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral