Tags

Type your tag names separated by a space and hit enter

Rapid detection of the most common high-risk human papillomaviruses by loop-mediated isothermal amplification.
J Virol Methods. 2011 Dec; 178(1-2):22-30.JV

Abstract

Persistent infection with high-risk human papillomavirus (HPV) is a major risk factor for development of cervical cancer. At present, polymerase chain reaction (PCR)-based methods, the most widely molecular tools used for HPV detection, are time-consuming and require expensive instruments. In this study, loop-mediated isothermal amplification (LAMP) was established for detection of HPV types 16, 18, 45 and 58 which are frequently found in Thailand. The optimal condition for detection of these high risk HPVs was 63°C for 60min. Since a white magnesium pyrophosphate precipitate is a characteristic by product of the LAMP reaction which can be visualized directly by the naked eye, the entire assay time of LAMP is 1h compared to 6-8h of for a nested PCR detection. The detection limit of LAMP assay was shown to be equivalent to nested PCR that could amplify 10(2) copies of HPV-18 and 10(3) copies of HPV 16, 45 and 58, as determined by either turbidity detection or agarose gel electrophoresis. No cross-reaction was observed, indicating that LAMP assay has high type-specificity. The assay showed successful detection of HPV in 56 clinical specimens. Using nested PCR as the gold standard, the sensitivity, specificity, negative predictive values and positive predictive values of LAMP assay were 100%. In conclusion, LAMP assay is a high efficiency, low cost diagnostic tool, useful for rapid, accurate, direct detection of HPV for clinical diagnosis.

Authors+Show Affiliations

Department of Clinical Chemistry, Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21903136

Citation

Saetiew, Chitladda, et al. "Rapid Detection of the Most Common High-risk Human Papillomaviruses By Loop-mediated Isothermal Amplification." Journal of Virological Methods, vol. 178, no. 1-2, 2011, pp. 22-30.
Saetiew C, Limpaiboon T, Jearanaikoon P, et al. Rapid detection of the most common high-risk human papillomaviruses by loop-mediated isothermal amplification. J Virol Methods. 2011;178(1-2):22-30.
Saetiew, C., Limpaiboon, T., Jearanaikoon, P., Daduang, S., Pientong, C., Kerdsin, A., & Daduang, J. (2011). Rapid detection of the most common high-risk human papillomaviruses by loop-mediated isothermal amplification. Journal of Virological Methods, 178(1-2), 22-30. https://doi.org/10.1016/j.jviromet.2011.08.007
Saetiew C, et al. Rapid Detection of the Most Common High-risk Human Papillomaviruses By Loop-mediated Isothermal Amplification. J Virol Methods. 2011;178(1-2):22-30. PubMed PMID: 21903136.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Rapid detection of the most common high-risk human papillomaviruses by loop-mediated isothermal amplification. AU - Saetiew,Chitladda, AU - Limpaiboon,Temduang, AU - Jearanaikoon,Patcharee, AU - Daduang,Sakda, AU - Pientong,Chamsai, AU - Kerdsin,Anusak, AU - Daduang,Jureerut, Y1 - 2011/08/27/ PY - 2011/01/15/received PY - 2011/08/04/revised PY - 2011/08/04/accepted PY - 2011/9/10/entrez PY - 2011/9/10/pubmed PY - 2012/2/9/medline SP - 22 EP - 30 JF - Journal of virological methods JO - J. Virol. Methods VL - 178 IS - 1-2 N2 - Persistent infection with high-risk human papillomavirus (HPV) is a major risk factor for development of cervical cancer. At present, polymerase chain reaction (PCR)-based methods, the most widely molecular tools used for HPV detection, are time-consuming and require expensive instruments. In this study, loop-mediated isothermal amplification (LAMP) was established for detection of HPV types 16, 18, 45 and 58 which are frequently found in Thailand. The optimal condition for detection of these high risk HPVs was 63°C for 60min. Since a white magnesium pyrophosphate precipitate is a characteristic by product of the LAMP reaction which can be visualized directly by the naked eye, the entire assay time of LAMP is 1h compared to 6-8h of for a nested PCR detection. The detection limit of LAMP assay was shown to be equivalent to nested PCR that could amplify 10(2) copies of HPV-18 and 10(3) copies of HPV 16, 45 and 58, as determined by either turbidity detection or agarose gel electrophoresis. No cross-reaction was observed, indicating that LAMP assay has high type-specificity. The assay showed successful detection of HPV in 56 clinical specimens. Using nested PCR as the gold standard, the sensitivity, specificity, negative predictive values and positive predictive values of LAMP assay were 100%. In conclusion, LAMP assay is a high efficiency, low cost diagnostic tool, useful for rapid, accurate, direct detection of HPV for clinical diagnosis. SN - 1879-0984 UR - https://www.unboundmedicine.com/medline/citation/21903136/Rapid_detection_of_the_most_common_high_risk_human_papillomaviruses_by_loop_mediated_isothermal_amplification_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0166-0934(11)00325-9 DB - PRIME DP - Unbound Medicine ER -