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Cited by 55Cervical Cancer and HPV ResearchReproductive tract infections researchBiosensors and Analytical Detection

Nonisotopic Detection and Typing of Human Papillomavirus DNA in Genital Samples by the Line Blot Assay

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Coutlée, François, Gravitt, Patti, Richardson, Harriet, Hankins, Catherine, Franco, Eduardo, Lapointe, Normand, Voyer, Hélène, The Canadian Women’s HIV Study Group·1999·Journal of Clinical Microbiology
DOI 10.1128/jcm.37.6.1852-1857.1999PMC PMC84968
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Abstract

The line blot assay, a gene amplification method that combines PCR with nonisotopic detection of amplified DNA, was evaluated for its ability to detect human papillomavirus (HPV) DNA in genital specimens. Processed samples were amplified with biotin-labeled primers for HPV detection (primers MY09, MY11, and HMB01) and for β-globin detection (primers PC03 and PC04). Amplified DNA products were hybridized by a reverse blot method with oligonucleotide probe mixtures fixed on a strip that allowed the identification of 27 HPV genotypes. The line blot assay was compared to a standard consensus PCR test in which HPV amplicons were detected with radiolabeled probes in a dot blot assay. Two hundred fifty-five cervicovaginal lavage specimens and cervical scrapings were tested in parallel by both PCR tests. The line blot assay consistently detected 25 copies of HPV type 18 per run. The overall positivity for the DNA of HPV types detectable by both methods was 37.7% (96 of 255 samples) by the line blot assay, whereas it was 43.5% (111 of 255 samples) by the standard consensus PCR assay. The sensitivity and specificity of the line blot assay reached 84.7% (94 of 111 samples) and 98.6% (142 of 144 samples), respectively. The agreement for HPV typing between the two PCR assays reached 83.9% (214 of 255 samples). Of the 37 samples with discrepant results, 33 (89%) were resolved by avoiding coamplification of β-globin and modifying the amplification parameters. With these modifications, the line blot assay compared favorably to an assay that used radiolabeled probes. Its convenience allows the faster analysis of samples for large-scale epidemiological studies. Also, the increased probe spectrum in this single hybridization assay permits more complete type discrimination.

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Crossref · 10.1128/jcm.37.6.1852-1857.1999

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  • Co-amplification of β-globin with HPV DNA or the presence of multiple HPV types can result in competition for assay reagents and hamper assay performance.

    “As previously reported with consensus PCR assays, the co-amplification of β-globin with HPV DNA or the presence of multiple HPV types can result in competition for assay reagents and hamper assay performance [21,22].”

    From Comparative evaluation of Allplex HPV28 and Anyplex II HPV28 assays for high-risk HPV genotyping in cervical samples (2025) · cites this paper as [21]

    The Results (page not available) report reduced HPV detection sensitivity with β-globin co-amplification and improved detection when it was avoided. The Discussion (page not available) says the line blot assay was less sensitive especially in samples with multiple HPV types. The paper does not establish competition for assay reagents as the mechanism for either effect. (LLM, conf=0.86)

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Cited by

Sorted by citations

Authors
Coutlée, F., Gravitt, P., Richardson, H., Hankins, C., Franco, E., Lapointe, N., Voyer, H., & The Canadian Women’s HIV Study Group
Affiliations
  • <!--label omitted: 1-->Départements de Microbiologie-Immunologie et de Pédiatrie, Université de Montréal,1
  • <!--label omitted: 3-->Roche Molecular Systems, Alameda, California3
  • McGill University (CA)
  • Sante Montreal (CA)
  • Centre Hospitalier Universitaire Sainte-Justine (CA)
  • <!--label omitted: 2-->Centre de Recherche et Département de Microbiologie et Infectiologie, Centre Hospitalier de l’Université de Montréal, Campus Notre-Dame,2
ORCID
Venue
Journal of Clinical Microbiology
Publisher
American Society for Microbiology
Published
1999-06
Volume
37
Issue
6
Pages
1852-1857
ISSN
  • 0095-1137 (ISSN-L)
  • 1098-660X
DOI
10.1128/jcm.37.6.1852-1857.1999
PMC
PMC84968
OpenAlex
W2158855921
Cited by
55
References
0
Citations by year
2025: 1 · 2022: 1 · 2021: 1 · 2017: 2 · 2013: 2 · 2012: 1
FWCI
3.29
Type
journal article
Language
en
Open access
Licence
journals.asm.org/non-commercial-tdm-license (tdm)
Locations
  • Journal of Clinical Microbiology (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Health Sciences › Medicine › Epidemiology › Cervical Cancer and HPV Research
  • Life Sciences › Immunology and Microbiology › Microbiology › Reproductive tract infections research
  • Physical Sciences › Engineering › Biomedical Engineering › Biosensors and Analytical Detection
Keywords
human papillomavirus, HPV testing, HPV genotyping, nonisotopic detection, biotin-labeled primers, cervical specimens, cervicovaginal lavage, assay sensitivity and specificity
MeSH terms
Cervix Uteri, DNA, Viral, Female, Genetic Techniques, HeLa Cells, Humans, Therapeutic Irrigation, Sensitivity and Specificity, Vagina, Vaginal Smears, Reproducibility of Results, Polymerase Chain Reaction, Papillomaviridae

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