Development and validation of a real-time PCR assay specifically detecting human papillomavirus 52 using the Roche LightCycler® 480 system

The recently released Linear Array® human papillomavirus (HPV) genotyping test (Roche Diagnostics) provides a standardized method for simultaneous detection of up to 37 individual HPV types. This test offers a rapid approach for detecting and longitudinal monitoring of patients infected with high-ri...

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Bibliographic Details
Published in:Journal of virological methods 2008-02, Vol.147 (2), p.290-296
Main Authors: Stevens, Matthew P., Garland, Suzanne M., Tabrizi, Sepehr N.
Format: Article
Language:English
Subjects:
Alphapapillomavirus - genetics
Alphapapillomavirus - isolation & purification
Base Sequence
Biological and medical sciences
Cervical cancer
Cervix Uteri - virology
Female
Fundamental and applied biological sciences. Psychology
Genotype
HPV type 52
Humans
LightCycler® 480
Microbiology
Molecular Sequence Data
Papillomavirus Infections - diagnosis
Papillomavirus Infections - virology
Polymerase Chain Reaction - methods
Real-time PCR
Sensitivity and Specificity
Sequence Alignment
Techniques used in virology
Uterine Cervical Diseases - diagnosis
Uterine Cervical Diseases - virology
Vaginal Smears
Virology
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Summary:The recently released Linear Array® human papillomavirus (HPV) genotyping test (Roche Diagnostics) provides a standardized method for simultaneous detection of up to 37 individual HPV types. This test offers a rapid approach for detecting and longitudinal monitoring of patients infected with high-risk (HR) HPV. However, it cannot rule out HPV52 infections in the presence of carcinogenic HPV genotypes 33, 35 and/or 58. As such, often only a non-definitive result of HPV52 presence can be reported. This study describes the development of a real-time PCR assay using an HPV52-specific hydrolysis probe in conjunction with the newly released Roche LightCycler® 480 system. HPV52 was readily detected among DNA extracts from samples previously identified with possible HPV52 by the linear array test. Specificity was analyzed using a panel of DNA extracts previously identified as containing single/multiple HPV types, with or without HPV52. This is a rapid and simple assay, which could be used as a supplementary test to the Linear Array, to verify the presence or absence of HPV52 among samples testing positive for either HPV33, 35, and/or 58. Laboratories already using the Linear Array HPV genotyping test could adopt this method once internally validated.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2007.09.018