CRISPR-based rapid and ultra-sensitive diagnostic test for Mycobacterium tuberculosis

被引:161
作者
Ai, Jing-Wen [1 ]
Zhou, Xian [1 ]
Xu, Teng [2 ]
Yang, Minling [2 ]
Chen, Yuanyuan [3 ]
He, Gui-Qing [4 ]
Pan, Ningp [4 ]
Cai, Yuwei [4 ]
Li, Yongjun [2 ]
Wang, Xiaorui [2 ]
Su, Hang [2 ]
Wang, Ting [5 ]
Zeng, Weiqi [2 ]
Zhang, Wen-Hong [1 ]
机构
[1] Fudan Univ, Huashan Hosp, Dept Infect Dis, 12 Wulumuqi Zhong Rd, Shanghai 200040, Peoples R China
[2] Vis Med Co Ltd, G10 BLDG,Huaxin Pk,31 Kefeng Ave, Guangzhou 510000, Guangdong, Peoples R China
[3] Hangzhou Red Cross Hosp, TB Diag & Treatment Ctr, Hangzhou, Zhejiang, Peoples R China
[4] Wenzhou Cent Hosp, Dept Infect Dis, Wenzhou, Peoples R China
[5] Jingan Dist Cent Hosp Shanghai, Dept Infect Dis, Shanghai, Peoples R China
关键词
CRISPR-MTB; tuberculosis; Mycobacterium tuberculosis complex (MTB); GeneXpert MTB; RIF; diagnosis; ADAPTIVE IMMUNITY; DNA;
D O I
10.1080/22221751.2019.1664939
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Rapid and simple-to-use diagnostic methods for tuberculosis are urgently needed. Recent development has unveiled the diagnostic power of the CRISPR system in the detection of viral infections. However, its potential use in detecting the Mycobacterium tuberculosis complex (MTB) remained unexplored. We developed a rapid CRISPR-based assay for TB detection and conducted a retrospective cohort study of 179 patients to evaluate the CRISPR-MTB test for identifying MTB in various forms of direct clinical samples. Its diagnostic performance was compared, in parallel with culture and the GeneXpert MTB/RIF assay (Xpert). The CRISPR-MTB test is highly sensitive with a near single-copy sensitivity, demands less sample input and offers shorter turnaround time than Xpert. When evaluated in the clinical cohort of both pulmonary and extra-pulmonary tuberculosis, the CRISPR-MTB test exhibited an overall improved sensitivity over both culture (79% vs 33%) and Xpert (79% vs 66%), without comprise in specificity (62/63, 98%). The CRISPR-MTB test exhibits an improved overall diagnostic performance over culture and Xpert across a variety of sample types, and offers great potential as a new diagnostic technique for both pulmonary and extra-pulmonary tuberculosis.
引用
收藏
页码:1361 / 1369
页数:9
相关论文
共 21 条
[1]   C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector [J].
Abudayyeh, Omar O. ;
Gootenberg, Jonathan S. ;
Konermann, Silvana ;
Joung, Julia ;
Slaymaker, Ian M. ;
Cox, David B. T. ;
Shmakov, Sergey ;
Makarova, Kira S. ;
Semenova, Ekaterina ;
Minakhin, Leonid ;
Severinov, Konstantin ;
Regev, Aviv ;
Lander, Eric S. ;
Koonin, Eugene V. ;
Zhang, Feng .
SCIENCE, 2016, 353 (6299)
[2]  
[Anonymous], 2013, Automated real-time nucleic acid amplification technology for rapid and simultaneous detection of tuberculosis and rifampicin resistance: Xpert MTB/RIF assay for the diagnosis of pulmonary and extrapulmonary TB in adults and children: policy update
[3]  
Association CM, 2005, CHIN CLIN TREATM GUI
[4]   CRISPR-Cas Systems: Prokaryotes Upgrade to Adaptive Immunity [J].
Barrangou, Rodolphe ;
Marraffini, Luciano A. .
MOLECULAR CELL, 2014, 54 (02) :234-244
[5]   Rapid Detection of Mycobacterium tuberculosis by Recombinase Polymerase Amplification [J].
Boyle, David S. ;
McNerney, Ruth ;
Low, Hwee Teng ;
Leader, Brandon Troy ;
Perez-Osorio, Ailyn C. ;
Meyer, Jessica C. ;
O'Sullivan, Denise M. ;
Brooks, David G. ;
Piepenburg, Olaf ;
Forrest, Matthew S. .
PLOS ONE, 2014, 9 (08)
[6]   DIAGNOSIS OF TUBERCULOSIS BY DNA AMPLIFICATION IN CLINICAL-PRACTICE EVALUATION [J].
BRISSONNOEL, A ;
AZNAR, C ;
CHUREAU, C ;
NGUYEN, S ;
PIERRE, C ;
BARTOLI, M ;
BONETE, R ;
PIALOUX, G ;
GICQUEL, B ;
GARRIGUE, G .
LANCET, 1991, 338 (8763) :364-366
[7]   CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity [J].
Chen, Janice S. ;
Ma, Enbo ;
Harrington, Lucas B. ;
Da Costa, Maria ;
Tian, Xinran ;
Palefsky, Joel M. ;
Doudna, Jennifer A. .
SCIENCE, 2018, 360 (6387) :436-+
[8]  
Dorman SE, 2018, LANCET INFECT DIS, V18, P76, DOI [10.1016/S1473-3099(17)30691-6, 10.1016/s1473-3099(17)30691-6]
[9]   Programmed DNA destruction by miniature CRISPR-Cas14 enzymes [J].
Harrington, Lucas B. ;
Burstein, David ;
Chen, Janice S. ;
Paez-Espino, David ;
Ma, Enbo ;
Witte, Isaac P. ;
Cofsky, Joshua C. ;
Kyrpides, Nikos C. ;
Banfield, Jillian F. ;
Doudna, Jennifer A. .
SCIENCE, 2018, 362 (6416) :839-+
[10]   A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity [J].
Jinek, Martin ;
Chylinski, Krzysztof ;
Fonfara, Ines ;
Hauer, Michael ;
Doudna, Jennifer A. ;
Charpentier, Emmanuelle .
SCIENCE, 2012, 337 (6096) :816-821