Tags

Type your tag names separated by a space and hit enter

Visual detection of high-risk human papillomavirus genotypes 16, 18, 45, 52, and 58 by loop-mediated isothermal amplification with hydroxynaphthol blue dye.
J Clin Microbiol. 2011 Oct; 49(10):3545-50.JC

Abstract

A simple, rapid, sensitive, qualitative, colorimetric loop-mediated isothermal amplification (LAMP) with hydroxynaphthol blue dye (HNB) was established to detect high-risk human papillomavirus (HPV) genotypes 16, 18, 45, 52, and 58. All initial validation studies with the control DNA proved to be type specific. The colorimetric type-specific LAMP assay could achieve a sensitivity of 10 to 100 copies at 63°C for 65 min, comparable to that of real-time PCR. In order to evaluate the reliability of HPV type-specific LAMP, the assay was further evaluated with HPV DNAs from a panel of 294 clinical specimens whose HPV status was previously determined with a novel one-step typing method with multiplex PCR. The tested panel comprised 108 HPV DNA-negative samples and 186 HPV-DNA-positive samples of 14 genotypes. The results showed that the sensitivity of HPV type-specific LAMP for HPV types 16, 18, 45, 52, and 58 was 100%, 100%, 100%, 100%, and 100%, respectively, and the specificity was 100%, 98.5%, 100%, 98.8%, and 99.2%, respectively, compared with a novel one-step typing method with multiplex PCR. No cross-reactivity with other HPV genotypes was observed. In conclusion, this qualitative and colorimetric LAMP assay has potential usefulness for the rapid screening of HPV genotype 16, 18, 45, 52, and 58 infections, especially in resource-limited hospitals or rural clinics of provincial and municipal regions in China.

Authors+Show Affiliations

State Key Laboratory for Molecular Virology and Genetic Engineering, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changbai Road 155, Changping District, Beijing 102206, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

21865423

Citation

Luo, Le, et al. "Visual Detection of High-risk Human Papillomavirus Genotypes 16, 18, 45, 52, and 58 By Loop-mediated Isothermal Amplification With Hydroxynaphthol Blue Dye." Journal of Clinical Microbiology, vol. 49, no. 10, 2011, pp. 3545-50.
Luo L, Nie K, Yang MJ, et al. Visual detection of high-risk human papillomavirus genotypes 16, 18, 45, 52, and 58 by loop-mediated isothermal amplification with hydroxynaphthol blue dye. J Clin Microbiol. 2011;49(10):3545-50.
Luo, L., Nie, K., Yang, M. J., Wang, M., Li, J., Zhang, C., Liu, H. T., & Ma, X. J. (2011). Visual detection of high-risk human papillomavirus genotypes 16, 18, 45, 52, and 58 by loop-mediated isothermal amplification with hydroxynaphthol blue dye. Journal of Clinical Microbiology, 49(10), 3545-50. https://doi.org/10.1128/JCM.00930-11
Luo L, et al. Visual Detection of High-risk Human Papillomavirus Genotypes 16, 18, 45, 52, and 58 By Loop-mediated Isothermal Amplification With Hydroxynaphthol Blue Dye. J Clin Microbiol. 2011;49(10):3545-50. PubMed PMID: 21865423.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Visual detection of high-risk human papillomavirus genotypes 16, 18, 45, 52, and 58 by loop-mediated isothermal amplification with hydroxynaphthol blue dye. AU - Luo,Le, AU - Nie,Kai, AU - Yang,Meng-Jie, AU - Wang,Miao, AU - Li,Jin, AU - Zhang,Chen, AU - Liu,Hong-Tu, AU - Ma,Xue-Jun, Y1 - 2011/08/24/ PY - 2011/8/26/entrez PY - 2011/8/26/pubmed PY - 2012/1/24/medline SP - 3545 EP - 50 JF - Journal of clinical microbiology JO - J. Clin. Microbiol. VL - 49 IS - 10 N2 - A simple, rapid, sensitive, qualitative, colorimetric loop-mediated isothermal amplification (LAMP) with hydroxynaphthol blue dye (HNB) was established to detect high-risk human papillomavirus (HPV) genotypes 16, 18, 45, 52, and 58. All initial validation studies with the control DNA proved to be type specific. The colorimetric type-specific LAMP assay could achieve a sensitivity of 10 to 100 copies at 63°C for 65 min, comparable to that of real-time PCR. In order to evaluate the reliability of HPV type-specific LAMP, the assay was further evaluated with HPV DNAs from a panel of 294 clinical specimens whose HPV status was previously determined with a novel one-step typing method with multiplex PCR. The tested panel comprised 108 HPV DNA-negative samples and 186 HPV-DNA-positive samples of 14 genotypes. The results showed that the sensitivity of HPV type-specific LAMP for HPV types 16, 18, 45, 52, and 58 was 100%, 100%, 100%, 100%, and 100%, respectively, and the specificity was 100%, 98.5%, 100%, 98.8%, and 99.2%, respectively, compared with a novel one-step typing method with multiplex PCR. No cross-reactivity with other HPV genotypes was observed. In conclusion, this qualitative and colorimetric LAMP assay has potential usefulness for the rapid screening of HPV genotype 16, 18, 45, 52, and 58 infections, especially in resource-limited hospitals or rural clinics of provincial and municipal regions in China. SN - 1098-660X UR - https://www.unboundmedicine.com/medline/citation/21865423/Visual_detection_of_high_risk_human_papillomavirus_genotypes_16_18_45_52_and_58_by_loop_mediated_isothermal_amplification_with_hydroxynaphthol_blue_dye_ L2 - http://jcm.asm.org/cgi/pmidlookup?view=long&pmid=21865423 DB - PRIME DP - Unbound Medicine ER -