Tags

Type your tag names separated by a space and hit enter

[Colorimetric detection of HPV6 and HPV16 by loop mediated isothermal amplification].
Bing Du Xue Bao. 2011 Jan; 27(1):64-70.BD

Abstract

A simple, rapid and sensitive colorimetric loop mediated isothermal amplification (LAMP) method was established to detect HPV6 and HPV 16 respectively. The method employed a set of four specially designed primers that recognized six distinct sequences of HPV6-E6 or HPV16-E7 for amplification of nucleic acid under isothermal conditions at 63 degrees C for one hour. The amplification process of LAMP was monitored by the addition of HNB (hydroxy naphthol blue) dye prior to amplification. A positive reaction was indicated by a color change from violet to sky blue and confirmed by real-time turbidimeter and agarose electrophoresis. Thirteen cervical swab samples having single infection with 13 different HPV genotypes were examined to evaluate the specificity. A serial dilution of a cloned plasmid containing HPV-E6 or HPV-E7 gene was examined to evaluate the sensitivity. The results showed that no cross-reaction with other HPV genotypes was observed. The colorimetric LAMP assay could achieve a sensitivity of 1000 copies, 10-20 times lower than that of real-time PCR. The assay was further evaluated with 62 clinical specimens and consistent results were obtained compared with the detection using Kai Pu HPV Genotyping Kit. We concluded that this colorimetric LAMP assay had potential usefulness for the rapid screening of the HPV6 or HPV16 infection in the laboratories and hospitals of provincial and municipal region in China.

Authors+Show Affiliations

Institute of Reproductive Immunology, Ji Nan University, Guangzhou 510632, China. tcblu@jnu.edu.cnNo 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

chi

PubMed ID

21462508

Citation

Lu, Chun-bin, et al. "[Colorimetric Detection of HPV6 and HPV16 By Loop Mediated Isothermal Amplification]." Bing Du Xue Bao = Chinese Journal of Virology, vol. 27, no. 1, 2011, pp. 64-70.
Lu CB, Luo L, Yang MJ, et al. [Colorimetric detection of HPV6 and HPV16 by loop mediated isothermal amplification]. Bing Du Xue Bao. 2011;27(1):64-70.
Lu, C. B., Luo, L., Yang, M. J., Nie, K., Wang, M., & Ma, X. J. (2011). [Colorimetric detection of HPV6 and HPV16 by loop mediated isothermal amplification]. Bing Du Xue Bao = Chinese Journal of Virology, 27(1), 64-70.
Lu CB, et al. [Colorimetric Detection of HPV6 and HPV16 By Loop Mediated Isothermal Amplification]. Bing Du Xue Bao. 2011;27(1):64-70. PubMed PMID: 21462508.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Colorimetric detection of HPV6 and HPV16 by loop mediated isothermal amplification]. AU - Lu,Chun-bin, AU - Luo,Le, AU - Yang,Meng-jie, AU - Nie,Kai, AU - Wang,Miao, AU - Ma,Xue-Jun, PY - 2011/4/6/entrez PY - 2011/4/6/pubmed PY - 2011/6/3/medline SP - 64 EP - 70 JF - Bing du xue bao = Chinese journal of virology JO - Bing Du Xue Bao VL - 27 IS - 1 N2 - A simple, rapid and sensitive colorimetric loop mediated isothermal amplification (LAMP) method was established to detect HPV6 and HPV 16 respectively. The method employed a set of four specially designed primers that recognized six distinct sequences of HPV6-E6 or HPV16-E7 for amplification of nucleic acid under isothermal conditions at 63 degrees C for one hour. The amplification process of LAMP was monitored by the addition of HNB (hydroxy naphthol blue) dye prior to amplification. A positive reaction was indicated by a color change from violet to sky blue and confirmed by real-time turbidimeter and agarose electrophoresis. Thirteen cervical swab samples having single infection with 13 different HPV genotypes were examined to evaluate the specificity. A serial dilution of a cloned plasmid containing HPV-E6 or HPV-E7 gene was examined to evaluate the sensitivity. The results showed that no cross-reaction with other HPV genotypes was observed. The colorimetric LAMP assay could achieve a sensitivity of 1000 copies, 10-20 times lower than that of real-time PCR. The assay was further evaluated with 62 clinical specimens and consistent results were obtained compared with the detection using Kai Pu HPV Genotyping Kit. We concluded that this colorimetric LAMP assay had potential usefulness for the rapid screening of the HPV6 or HPV16 infection in the laboratories and hospitals of provincial and municipal region in China. SN - 1000-8721 UR - https://www.unboundmedicine.com/medline/citation/21462508/[Colorimetric_detection_of_HPV6_and_HPV16_by_loop_mediated_isothermal_amplification]_ DB - PRIME DP - Unbound Medicine ER -