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	Rapid colorimetric testing for pyrazinamide susceptibility of m. tuberculosis by a pcr-based in-vitro synthesized pyrazinamidase method
	Zhou, Man1,3; Geng, Xuelei1,3; Chen, Jun2; Wang, Xude1; Wang, Dianbing1; Deng, Jiaoyu1; Zhang, Zhiping1; Wang, Weihua2; Zhang, Xian-En1; Wei, Hongping1
刊名	Plos one
	2011-11-10
卷号	6期号:11页码:7
ISSN号	1932-6203
DOI	10.1371/journal.pone.0027654
通讯作者	Zhou, man()
英文摘要	Pyrazinamide (pza) is an important first-line anti-tuberculosis drug. but pza susceptibility test is challenging because pza activity is optimal only in an acid environment that inhibits the growth of m. tuberculosis. for current phenotypic methods, inconsistent results between different labs have been reported. direct sequencing of pnca gene is being considered as an accurate predictor for pza susceptibility, but this approach needs expensive sequencers and a mutation database to report the results. an in-vitro synthesized pyrazinamidase (pzase) assay was developed based on pcr amplification of pnca gene and an in vitro wheat germ system to express the pnca gene into pzase. the activity of the synthesized pzase was used as an indicator for pza susceptibility. fifty-one clinical isolates were tested along with pnca sequencing and the bactec mgit 960 methods. the in-vitro synthesized pzase assay was able to detect pza susceptibility of m. tuberculosis within 24 h through observing the color difference either by a spectrometer or naked eyes. this method showed agreements of 100% (33/33) and 88% (14/16) with the pnca sequencing method, and agreements of 96% (27/28) and 65% (15/23) with the bactec mgit 960 method, for susceptible and resistant strains, respectively. the novel in-vitro synthesized pzase assay has significant advantages over current methods, such as its fast speed, simplicity, no need for expensive equipment, and the potentials of being a direct test, predicting resistance level and easy reading results by naked eyes. after confirmation by more clinical tests, this method may provide a radical change to the current pza susceptibility assays.
WOS关键词	RESISTANT MYCOBACTERIUM-TUBERCULOSIS ; PNCA MUTATIONS ; MAJOR MECHANISM ; GENE ; SYSTEM ; STRAINS ; REGION
WOS研究方向	Science & Technology - Other Topics
WOS类目	Multidisciplinary Sciences
语种	英语
出版者	PUBLIC LIBRARY SCIENCE
WOS记录号	WOS:000297348800040
内容类型	期刊论文
URI标识	http://www.corc.org.cn/handle/1471x/2375920
专题	武汉病毒研究所
通讯作者	Zhou, Man
作者单位	1.Chinese Acad Sci, State Key Lab Virol, Wuhan Inst Virol, Wuhan, Peoples R China 2.Wuan TB Control Ctr, Wuhan, Peoples R China 3.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
推荐引用方式 GB/T 7714	Zhou, Man,Geng, Xuelei,Chen, Jun,et al. Rapid colorimetric testing for pyrazinamide susceptibility of m. tuberculosis by a pcr-based in-vitro synthesized pyrazinamidase method[J]. Plos one,2011,6(11):7.
APA	Zhou, Man.,Geng, Xuelei.,Chen, Jun.,Wang, Xude.,Wang, Dianbing.,...&Wei, Hongping.(2011).Rapid colorimetric testing for pyrazinamide susceptibility of m. tuberculosis by a pcr-based in-vitro synthesized pyrazinamidase method.Plos one,6(11),7.
MLA	Zhou, Man,et al."Rapid colorimetric testing for pyrazinamide susceptibility of m. tuberculosis by a pcr-based in-vitro synthesized pyrazinamidase method".Plos one 6.11(2011):7.

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