HPV DNA Detection and Typing in Cervical Scrapes

Snijders, Peter J. F.; van den Brule, Adriaan J. C.; Jacobs, Marcel V.; Pol, Renea P.; Meijer, Chris J. L. M.

doi:10.1385/1-59259-982-6:101

Cited by 63Cervical Cancer and HPV ResearchReproductive tract infections researchHead and Neck Cancer Studies

HPV DNA Detection and Typing in Cervical Scrapes

Snijders, Peter J. F., van den Brule, Adriaan J. C., Jacobs, Marcel V., Pol, Renea P., Meijer, Chris J. L. M.·2005·Human Papillomaviruses

DOI 10.1385/1-59259-982-6:101

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Abstract

Polymerase chain reaction (PCR)-based assays that use consensus primers to detect DNA of a broad spectrum of human papillomavirus (HPV) types in a single assay belong to the most frequently used methods to detect HPV in clinical specimens. Here, we describe in detail one of these assays, the so-called GP5+/6+ PCR method, which can be used to detect and type HPV DNA in crude extracts of cervical scrapes and biopsy specimens. Following PCR with GP5+ and GP6+ primers, the latter of which is biotinylated at its 5' end, the presence of DNA of any of the high-risk genotypes can easily be determined by an enzyme immunoassay (EIA). In this assay, PCR products are captured in streptavidin-coated wells of a microtiter plate, denatured by alkaline treatment, and hybridized to cocktails of digoxigenin-labeled oligonucleotides specific for high-risk or low-risk HPV types. The resulting hybrids can then be detected by alkaline phosphatase conjugated anti-digoxigenin polyclonal antibodies and substrate followed by optical density reading. Subsequently, EIA-positive PCR products can be typed by a reverse line blot genotyping procedure that, using a miniblotter device, enables typing of up to 39 samples with specific oligonucleotide probes for 37 different HPV (sub)types in a single assay.

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Anyplex II HPV HR Detection was performed blinded for earlier obtained GP5+/6+-PCR-EIA results and cyto-/histopathology outcomes, and data were correlated afterwards.
“Anyplex TM II HPV HR Detection was performed blinded for earlier obtained GP5+/6+-PCR-EIA results [5] and cyto-/histopathology outcomes, and data were correlated afterwards.”
From Clinical validation of Anyplex™ II HPV HR Detection according to the guidelines for HPV test requirements for cervical cancer screening (2016) · cites this paper as [5]

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Detecting and Typing Human Papillomaviruses By PCR Enzyme-immunoassay
Jun, W. (2000)

Authors: Snijders, P. J. F., van den Brule, A. J. C., Jacobs, M. V., Pol, R. P., & Meijer, C. J. L. M.
Affiliations: Amsterdam UMC Location Vrije Universiteit Amsterdam (NL)
ORCID: Snijders, Peter J. F.
Meijer, Chris J. L. M.
Venue: Human Papillomaviruses
Publisher: Humana Press
Published: 2005-08-08
Volume: 119
Pages: 101-114
DOI: 10.1385/1-59259-982-6:101
OpenAlex: W83911031
Cited by: 62
References: 12
Citations by year: 2025: 1 · 2023: 1 · 2022: 1 · 2021: 1 · 2020: 2 · 2019: 2 · 2018: 1 · 2017: 2 · 2016: 2 · 2015: 1 · 2014: 2 · 2013: 4 · 2012: 13
FWCI: 12.32
Type: book chapter
Language: en
Open access: closed
Locations: Humana Press eBooks (publishedVersion)
PubMed (publishedVersion)
Topics: Health Sciences › Medicine › Epidemiology › Cervical Cancer and HPV Research
Life Sciences › Immunology and Microbiology › Microbiology › Reproductive tract infections research
Health Sciences › Medicine › Otorhinolaryngology › Head and Neck Cancer Studies
Keywords: human papillomavirus, HPV testing, HPV genotyping, cervical scrapes, cervical biopsy, high-risk HPV genotypes, reverse line blot, enzyme immunoassay, alkaline phosphatase detection, consensus primers
MeSH terms: Base Sequence, Cervix Uteri, DNA, Viral, Female, Globins, Humans, Nucleic Acid Hybridization, Sensitivity and Specificity, Polymerase Chain Reaction, DNA Primers, Papillomaviridae

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