ArticleMolecular pathology : MP2001
Rapid real time PCR to distinguish between high risk human papillomavirus types 16 and 18.
Article in Molecular pathology : MP, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 15 papers.
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15 citing papers in PubMed, 44 citations in OpenAlex.
- Identification of HPV16-p16Epigenetics · 2021Article
- One step DNA amplification of mammalian cells in picoliter microwell arrays.RSC advances · 2019Article
- HPV infection and pre-term birth: a data-linkage study using Scottish Health Data.Wellcome open research · 2019Article
- Detection of human papillomavirus (HPV) DNA prevalence and p53 codon 72 (Arg72Pro) polymorphism in prostate cancer in a Greek group of patients.Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine · 2014Article
- From Human Papillomavirus (HPV) Detection to Cervical Cancer Prevention in Clinical Practice.Cancers · 2014Article
- Comparing the Performance of Hybrid Capture II and Polymerase Chain Reaction (PCR) for the Identification of Cervical Dysplasia in the Screening and Diagnostic Settings.Clinical Medicine Insights. Oncology · 2013Article
- Article
- Quantitative human papillomavirus 16 and 18 levels in incident infections and cervical lesion development.Journal of medical virology · 2009Article
- Loop-mediated isothermal amplification method for detection of human papillomavirus type 6, 11, 16, and 18.Journal of medical virology · 2007Article
- Human papilloma virus in squamous carcinoma of the head and neck: a study of cases in south east Scotland.Journal of clinical pathology · 2007Article
- Persistent high risk HPV infection associated with development of cervical neoplasia in a prospective population study.Journal of clinical pathology · 2005Article
- Advances in real-time PCR: application to clinical laboratory diagnostics.Advances in clinical chemistry · 2005Review
- Multiple high risk HPV infections are common in cervical neoplasia and young women in a cervical screening population.Journal of clinical pathology · 2004Article
- The causal relation between human papillomavirus and cervical cancer.Journal of clinical pathology · 2002Review
- LightCycler multiplex PCR for the laboratory diagnosis of common viral infections of the central nervous system.Journal of clinical microbiology · 2001Article
Corrections and comments
- Erratum issued
Authors and funding
7 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
aimsTo assess the validity and practicality of real time polymerase chain reaction (PCR) for human papillomavirus (HPV) testing in combination with liquid based cytology samples for cervical screening.
methodsReal time PCR using consensus (GPS+/6+) and type specific primers was developed to detect genital HPV types. This provides rapid, efficient amplification followed by denaturation of the product and computer analysis of the kinetics data that are generated. Liquid based cytology samples were obtained from patients attending routine cervical screening clinics. DNA was extracted from the residual cellular suspension after cytology using spin columns.
resultsReal time PCR successfully distinguished between HPV-16 and HPV-18 on the basis of amplification with consensus primers followed by DNA melting temperature (Tm) analysis. Sensitivities of one to 10 copies of HPV-16 (mean Tm = 79.4 degrees C; 2 SD, 0.8) and four to 40 copies of HPV-18 (mean Tm = 80.4 degrees C; 2 SD, 0.4) were obtained. In a mixed population of SiHa and HeLa cells containing known copy numbers of HPV-16 and HPV-18 genomes, HPV-16 and HPV-18 products were clearly separated by Tm analysis in mixtures varying from equivalence to 111000. Together with detailed melt analysis, type specific primers from the same region of the L1 gene confirmed the differential ability of this system. The method was applied to 100 liquid based cytology samples where HPV status using conventional GP5+/6+ PCR was already known. There was 95% agreement between the methods, with 55 positives detected by conventional PCR and 59 with real time PCR. The method was then tested on 200 routine liquid based cytology samples. Approximately 10% were positive by real time PCR, most of which were classified as HPV-16 by detailed melt analysis. Thirteen (6.8%) HPV positives were identified in 189 samples showing no evidence of cervical cytological abnormality.
conclusionsReal time PCR is a rapid, efficient method for the detection of HPV with the separation of HPV-16 and HPV-18 on the basis of differential Tm. Preliminary results suggest it could prove
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