Abstract:
:mRNAs of human papillomaviruses (HPV) 16 and 18 were detected in cancer-derived cell lines and genital tract biopsy specimens by a novel hybridization assay. Biotinylated whole genomic HPV DNA probes were hybridized in solution to extracted total nucleic acids. Hybrids between the labeled probes and RNA transcripts were captured on a microplate coated with an antibiotin antibody. Bound hybrids were incubated with a beta-galactosidase-labeled monoclonal antibody to DNA-RNA hybrids and measured by the addition of a fluorogenic substrate. HPV 18 and HPV 16 mRNAs were detected in nucleic acids from 2.3 x 10(3) HeLa cells and 10(4) SiHa cells, respectively. The specificity of the assay for mRNA was demonstrated by the low reactivity of nucleic acids from SiHa cells after treatment with T1 RNase and by the selective reactivity of cellular nucleic acids which bound to an oligo(dT) column. With HPV 16 subgenomic probes, E6-E7 transcripts but not L1-L2 transcripts were detected in SiHa cells. Tests of 58 biopsy specimens from 31 patients showed that the detection of HPV 16 and HPV 18 transcripts in tissue specimens was feasible. Analysis of biopsy specimens with subgenomic probes revealed HPV 16 E6-E7 transcripts in all specimens that reacted with the whole genomic probe, while L1-L2 transcripts were found infrequently.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Coutlée F,Shah KV,Rader JS,Currie JL,Viscidi RPdoi
10.1128/JCM.29.5.968-974.1991keywords:
subject
Has Abstractpub_date
1991-05-01 00:00:00pages
968-74issue
5eissn
0095-1137issn
1098-660Xjournal_volume
29pub_type
杂志文章abstract::Five serological assays were evaluated for the diagnosis of brucellosis in goats: the rose bengal test (RBT), complement fixation test (CFT), radial immunodiffusion (RID) with Brucella and Yersinia enterocolitica O:9 polysaccharides, counterimmunoelectrophoresis (CIEP) with cytosol, and enzyme-linked immunosorbent ass...
journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
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pub_type: 杂志文章,多中心研究
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.28.7.1586-1590.1990
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.18.3.495-499.1983
更新日期:1983-09-01 00:00:00
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.34.11.2713-2717.1996
更新日期:1996-11-01 00:00:00
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.12.3.462-468.1980
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journal_title:Journal of clinical microbiology
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更新日期:2003-11-01 00:00:00
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journal_title:Journal of clinical microbiology
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更新日期:2007-07-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.7.2153-2156.1998
更新日期:1998-07-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章,多中心研究
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更新日期:2008-08-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00676-08
更新日期:2008-08-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.13.1.184-194.1981
更新日期:1981-01-01 00:00:00
abstract::Two previously demonstrated sensitive and specific enzyme immunoassays (EIAs) for rotavirus, one using polyclonal and monoclonal antisera (TestPack Rotavirus [TPK]; Abbott Laboratories) and the other using only monoclonal anti-rotavirus antibodies (Rotaclone [RTC]; Cambridge BioScience Corporation), were evaluated as ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.28.6.1280-1283.1990
更新日期:1990-06-01 00:00:00