Application of hyperbranched rolling circle amplification for direct detection of mycobacterium tuberculosis in clinical sputum specimens.

<h4>Background</h4>Global tuberculosis (TB) control is encumbered by the lack of a rapid and simple detection method for diagnosis, especially in low-resource areas. An isothermal amplification method, hyperbranched rolling circle amplification (HRCA), was optimized to detect Mycobacteri...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Yang Liu, Yan-Ling Guo, Guang-Lu Jiang, Shi-Jie Zhou, Qi Sun, Xi Chen, Xiu-Jun Chang, Ai-Ying Xing, Feng-Jiao Du, Hong-Yan Jia, Zong-De Zhang
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2b6d92a673d8407a8ec98756343adea1
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:2b6d92a673d8407a8ec98756343adea1
record_format	dspace
spelling	oai:doaj.org-article:2b6d92a673d8407a8ec98756343adea12021-11-18T07:43:08ZApplication of hyperbranched rolling circle amplification for direct detection of mycobacterium tuberculosis in clinical sputum specimens.1932-620310.1371/journal.pone.0064583https://doaj.org/article/2b6d92a673d8407a8ec98756343adea12013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23750210/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Global tuberculosis (TB) control is encumbered by the lack of a rapid and simple detection method for diagnosis, especially in low-resource areas. An isothermal amplification method, hyperbranched rolling circle amplification (HRCA), was optimized to detect Mycobacterium tuberculosis (Mtb) in clinical sputum specimens.<h4>Methods</h4>A clinical validation study was performed to assess the diagnostic accuracy of HRCA. In order to analyze the detection limit of HRCA under optimal conditions, the method was initially used to detect purified H37Rv strain DNA and culture suspensions. Next, three strains of Mycobacterium tuberculosis complex (MTC) and eight strains of non-tuberculosis mycobacterium (NTM) were analyzed in order to evaluate specificity. Sputum specimens from 136 patients with diagnosed pulmonary TB, 38 lung cancer patients, and 34 healthy donors were tested by HRCA to validate the clinical application of HRCA for the rapid detection of Mtb.<h4>Results</h4>The detection limit of HRCA for purified H37Rv DNA and culture suspensions was 740 aM and 200cfu/ml, respectively. The results of all MTC strains were positive in contrast to the NTM specimens which were all negative. The detection sensitivity for the 136 sputum specimens from TB patients was 77.2% (105/136), which was slightly lower than that of quantitative real-time PCR(79.4%, 108/136) and culture (80.9%,110/136). The sensitivity of all three methods was statistically higher than smear microscopy (44.9%, 61/136). The overall specificity of HRCA was 98.6% (71/72) which was similar to that of quantitative real-time PCR (qRT-PCR) and smear/culture methods (100%, 72/72).<h4>Conclusions</h4>Use of the HRCA assay for detection of Mtb within clinical sputum specimens was demonstrated to be highly sensitive and specific. Moreover, the performance of HRCA is simple and cost-effective compared with qRT-PCR and is less time consuming than culture. Therefore, HRCA is a promising TB diagnostic tool that can be used routinely in low-resource clinical settings.Yang LiuYan-Ling GuoGuang-Lu JiangShi-Jie ZhouQi SunXi ChenXiu-Jun ChangAi-Ying XingFeng-Jiao DuHong-Yan JiaZong-De ZhangPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 6, p e64583 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yang Liu Yan-Ling Guo Guang-Lu Jiang Shi-Jie Zhou Qi Sun Xi Chen Xiu-Jun Chang Ai-Ying Xing Feng-Jiao Du Hong-Yan Jia Zong-De Zhang Application of hyperbranched rolling circle amplification for direct detection of mycobacterium tuberculosis in clinical sputum specimens.
description	<h4>Background</h4>Global tuberculosis (TB) control is encumbered by the lack of a rapid and simple detection method for diagnosis, especially in low-resource areas. An isothermal amplification method, hyperbranched rolling circle amplification (HRCA), was optimized to detect Mycobacterium tuberculosis (Mtb) in clinical sputum specimens.<h4>Methods</h4>A clinical validation study was performed to assess the diagnostic accuracy of HRCA. In order to analyze the detection limit of HRCA under optimal conditions, the method was initially used to detect purified H37Rv strain DNA and culture suspensions. Next, three strains of Mycobacterium tuberculosis complex (MTC) and eight strains of non-tuberculosis mycobacterium (NTM) were analyzed in order to evaluate specificity. Sputum specimens from 136 patients with diagnosed pulmonary TB, 38 lung cancer patients, and 34 healthy donors were tested by HRCA to validate the clinical application of HRCA for the rapid detection of Mtb.<h4>Results</h4>The detection limit of HRCA for purified H37Rv DNA and culture suspensions was 740 aM and 200cfu/ml, respectively. The results of all MTC strains were positive in contrast to the NTM specimens which were all negative. The detection sensitivity for the 136 sputum specimens from TB patients was 77.2% (105/136), which was slightly lower than that of quantitative real-time PCR(79.4%, 108/136) and culture (80.9%,110/136). The sensitivity of all three methods was statistically higher than smear microscopy (44.9%, 61/136). The overall specificity of HRCA was 98.6% (71/72) which was similar to that of quantitative real-time PCR (qRT-PCR) and smear/culture methods (100%, 72/72).<h4>Conclusions</h4>Use of the HRCA assay for detection of Mtb within clinical sputum specimens was demonstrated to be highly sensitive and specific. Moreover, the performance of HRCA is simple and cost-effective compared with qRT-PCR and is less time consuming than culture. Therefore, HRCA is a promising TB diagnostic tool that can be used routinely in low-resource clinical settings.
format	article
author	Yang Liu Yan-Ling Guo Guang-Lu Jiang Shi-Jie Zhou Qi Sun Xi Chen Xiu-Jun Chang Ai-Ying Xing Feng-Jiao Du Hong-Yan Jia Zong-De Zhang
author_facet	Yang Liu Yan-Ling Guo Guang-Lu Jiang Shi-Jie Zhou Qi Sun Xi Chen Xiu-Jun Chang Ai-Ying Xing Feng-Jiao Du Hong-Yan Jia Zong-De Zhang
author_sort	Yang Liu
title	Application of hyperbranched rolling circle amplification for direct detection of mycobacterium tuberculosis in clinical sputum specimens.
title_short	Application of hyperbranched rolling circle amplification for direct detection of mycobacterium tuberculosis in clinical sputum specimens.
title_full	Application of hyperbranched rolling circle amplification for direct detection of mycobacterium tuberculosis in clinical sputum specimens.
title_fullStr	Application of hyperbranched rolling circle amplification for direct detection of mycobacterium tuberculosis in clinical sputum specimens.
title_full_unstemmed	Application of hyperbranched rolling circle amplification for direct detection of mycobacterium tuberculosis in clinical sputum specimens.
title_sort	application of hyperbranched rolling circle amplification for direct detection of mycobacterium tuberculosis in clinical sputum specimens.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/2b6d92a673d8407a8ec98756343adea1
work_keys_str_mv	AT yangliu applicationofhyperbranchedrollingcircleamplificationfordirectdetectionofmycobacteriumtuberculosisinclinicalsputumspecimens AT yanlingguo applicationofhyperbranchedrollingcircleamplificationfordirectdetectionofmycobacteriumtuberculosisinclinicalsputumspecimens AT guanglujiang applicationofhyperbranchedrollingcircleamplificationfordirectdetectionofmycobacteriumtuberculosisinclinicalsputumspecimens AT shijiezhou applicationofhyperbranchedrollingcircleamplificationfordirectdetectionofmycobacteriumtuberculosisinclinicalsputumspecimens AT qisun applicationofhyperbranchedrollingcircleamplificationfordirectdetectionofmycobacteriumtuberculosisinclinicalsputumspecimens AT xichen applicationofhyperbranchedrollingcircleamplificationfordirectdetectionofmycobacteriumtuberculosisinclinicalsputumspecimens AT xiujunchang applicationofhyperbranchedrollingcircleamplificationfordirectdetectionofmycobacteriumtuberculosisinclinicalsputumspecimens AT aiyingxing applicationofhyperbranchedrollingcircleamplificationfordirectdetectionofmycobacteriumtuberculosisinclinicalsputumspecimens AT fengjiaodu applicationofhyperbranchedrollingcircleamplificationfordirectdetectionofmycobacteriumtuberculosisinclinicalsputumspecimens AT hongyanjia applicationofhyperbranchedrollingcircleamplificationfordirectdetectionofmycobacteriumtuberculosisinclinicalsputumspecimens AT zongdezhang applicationofhyperbranchedrollingcircleamplificationfordirectdetectionofmycobacteriumtuberculosisinclinicalsputumspecimens
_version_	1718423025879089152

Application of hyperbranched rolling circle amplification for direct detection of mycobacterium tuberculosis in clinical sputum specimens.

Ejemplares similares