Detection of human papillomavirus type-16 DNA utilising microtitre-plate based amplification reactions and a solid-phase enzyme-immunoassay detection system

S Cavuslu; W G Starkey; J N Kaye; C Biswas; C Mant; B Kell; P Rice; J M Best; J Cason

doi:10.1016/0166-0934(95)01988-x

Detection of human papillomavirus type-16 DNA utilising microtitre-plate based amplification reactions and a solid-phase enzyme-immunoassay detection system

S Cavuslu, W G Starkey, J N Kaye, C Biswas, C Mant, B Kell, P Rice, J M Best, J Cason

King's College London

Richard Dimbleby Laboratory of Cancer Virology, Department of Virology, Rayne Institute, United Medical and Dental School of Guys, St Thomas' Hospital, London, U.K.

Research output: Contribution to journal › Article › peer-review

16 Citations (Scopus)

Abstract

The development of a nested polymerase chain reaction (PCR) assay to detect low concentrations of human papillomavirus type-16 (HPV-16) DNA for epidemiological studies is described. The PCR utilises primers located in the E5 open reading frame, has an analytical sensitivity of 4 HPV-16 genomes and does not produce amplicons from other common genital HPVs (types-6, -11, -18, -31 and 33). This assay was carried out in 96-well plates utilising internal primers labelled with dinitrophenol (DNP) and biotin so that amplicons can be captured onto streptavidincoated plates and detected using an alkaline phosphatase-labelled monoclonal antibody to DNP. The assay was effective for detecting HPV-16 DNA in plasmids, cell-lines and, both freshly collected or archival (formalin-fixed/paraffin embedded) clinical specimens. This system is therefore suitable for epidemiological studies to identify individuals infected with HPV-16 DNA in episomal form who may be at increased risk of developing anogenital carcinomas.

Original language	English
Pages (from-to)	59-69
Number of pages	11
Journal	Journal of Virological Methods
Volume	58
Issue number	1-2
DOIs	https://doi.org/10.1016/0166-0934(95)01988-x
Publication status	Published - 26 Apr 1996

Keywords

Cervical Intraepithelial Neoplasia/pathology
DNA Primers
DNA, Viral/analysis
Evaluation Studies as Topic
Female
HeLa Cells
Humans
Immunoenzyme Techniques
Oncogene Proteins, Viral/genetics
Papillomaviridae/genetics
Papillomavirus E7 Proteins
Polymerase Chain Reaction/methods
Repressor Proteins
Sensitivity and Specificity
Tumor Cells, Cultured
Uterine Cervical Neoplasms/pathology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/0166-0934(95)01988-x

Cite this

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title = "Detection of human papillomavirus type-16 DNA utilising microtitre-plate based amplification reactions and a solid-phase enzyme-immunoassay detection system",

abstract = "The development of a nested polymerase chain reaction (PCR) assay to detect low concentrations of human papillomavirus type-16 (HPV-16) DNA for epidemiological studies is described. The PCR utilises primers located in the E5 open reading frame, has an analytical sensitivity of 4 HPV-16 genomes and does not produce amplicons from other common genital HPVs (types-6, -11, -18, -31 and 33). This assay was carried out in 96-well plates utilising internal primers labelled with dinitrophenol (DNP) and biotin so that amplicons can be captured onto streptavidincoated plates and detected using an alkaline phosphatase-labelled monoclonal antibody to DNP. The assay was effective for detecting HPV-16 DNA in plasmids, cell-lines and, both freshly collected or archival (formalin-fixed/paraffin embedded) clinical specimens. This system is therefore suitable for epidemiological studies to identify individuals infected with HPV-16 DNA in episomal form who may be at increased risk of developing anogenital carcinomas.",

keywords = "Cervical Intraepithelial Neoplasia/pathology, DNA Primers, DNA, Viral/analysis, Evaluation Studies as Topic, Female, HeLa Cells, Humans, Immunoenzyme Techniques, Oncogene Proteins, Viral/genetics, Papillomaviridae/genetics, Papillomavirus E7 Proteins, Polymerase Chain Reaction/methods, Repressor Proteins, Sensitivity and Specificity, Tumor Cells, Cultured, Uterine Cervical Neoplasms/pathology",

author = "S Cavuslu and Starkey, {W G} and Kaye, {J N} and C Biswas and C Mant and B Kell and P Rice and Best, {J M} and J Cason",

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doi = "10.1016/0166-0934(95)01988-x",

language = "English",

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AU - Cavuslu, S

AU - Starkey, W G

AU - Kaye, J N

AU - Biswas, C

AU - Mant, C

AU - Kell, B

AU - Rice, P

AU - Best, J M

AU - Cason, J

PY - 1996/4/26

Y1 - 1996/4/26

N2 - The development of a nested polymerase chain reaction (PCR) assay to detect low concentrations of human papillomavirus type-16 (HPV-16) DNA for epidemiological studies is described. The PCR utilises primers located in the E5 open reading frame, has an analytical sensitivity of 4 HPV-16 genomes and does not produce amplicons from other common genital HPVs (types-6, -11, -18, -31 and 33). This assay was carried out in 96-well plates utilising internal primers labelled with dinitrophenol (DNP) and biotin so that amplicons can be captured onto streptavidincoated plates and detected using an alkaline phosphatase-labelled monoclonal antibody to DNP. The assay was effective for detecting HPV-16 DNA in plasmids, cell-lines and, both freshly collected or archival (formalin-fixed/paraffin embedded) clinical specimens. This system is therefore suitable for epidemiological studies to identify individuals infected with HPV-16 DNA in episomal form who may be at increased risk of developing anogenital carcinomas.

AB - The development of a nested polymerase chain reaction (PCR) assay to detect low concentrations of human papillomavirus type-16 (HPV-16) DNA for epidemiological studies is described. The PCR utilises primers located in the E5 open reading frame, has an analytical sensitivity of 4 HPV-16 genomes and does not produce amplicons from other common genital HPVs (types-6, -11, -18, -31 and 33). This assay was carried out in 96-well plates utilising internal primers labelled with dinitrophenol (DNP) and biotin so that amplicons can be captured onto streptavidincoated plates and detected using an alkaline phosphatase-labelled monoclonal antibody to DNP. The assay was effective for detecting HPV-16 DNA in plasmids, cell-lines and, both freshly collected or archival (formalin-fixed/paraffin embedded) clinical specimens. This system is therefore suitable for epidemiological studies to identify individuals infected with HPV-16 DNA in episomal form who may be at increased risk of developing anogenital carcinomas.

KW - Cervical Intraepithelial Neoplasia/pathology

KW - DNA Primers

KW - DNA, Viral/analysis

KW - Evaluation Studies as Topic

KW - Female

KW - HeLa Cells

KW - Humans

KW - Immunoenzyme Techniques

KW - Oncogene Proteins, Viral/genetics

KW - Papillomaviridae/genetics

KW - Papillomavirus E7 Proteins

KW - Polymerase Chain Reaction/methods

KW - Repressor Proteins

KW - Sensitivity and Specificity

KW - Tumor Cells, Cultured

KW - Uterine Cervical Neoplasms/pathology

U2 - 10.1016/0166-0934(95)01988-x

DO - 10.1016/0166-0934(95)01988-x

M3 - Article

C2 - 8783151

SN - 0166-0934

VL - 58

SP - 59

EP - 69

JO - Journal of Virological Methods

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IS - 1-2

ER -

Detection of human papillomavirus type-16 DNA utilising microtitre-plate based amplification reactions and a solid-phase enzyme-immunoassay detection system

Abstract

Keywords

UN SDGs

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