TB-QUICK: CRISPR-Cas12b-assisted rapid and sensitive detection of Mycobacterium tuberculosis

被引：67

作者：

Sam, I. Kuan ^{[1
]}

Chen, Ying-Ying ^{[1
]}

Ma, Jun ^{[2
]}

Li, Shi-Yuan ^{[3
]}

Ying, Ruo-Yan ^{[2
]}

Li, Lin-Xian ^{[4
]}

Ji, Ping ^{[1
]}

Wang, Shu-Jun ^{[1
]}

Xu, Jie ^{[5
]}

Bao, Yu-Jie ^{[5
]}

Zhao, Guo-Ping ^{[6
,7
]}

Zheng, Hua-Jun ^{[7
]}

Wang, Jin ^{[4
]}

Sha, Wei ^{[2
]}

Wang, Ying ^{[1
,7
,8
]}

机构：

[1] Shanghai Jiao Tong Univ, Shanghai Inst Immunol, Dept Microbiol & Immunol, Sch Med, Shanghai 200025, Peoples R China

[2] Tongji Univ, Shanghai Pulm Hosp, Clin & Res Ctr TB, Shanghai Key Lab TB,Sch Med, Shanghai 200433, Peoples R China

[3] Shanghai TOLO Biotechnol Co Ltd, Shanghai 200233, Peoples R China

[4] Shenzhen Univ, Shenzhen Peoples Hosp 2, Shenzhen Inst Translat Med, Dept Clin Lab,Affiliated Hosp 1, Shenzhen 518035, Peoples R China

[5] Shanghai Jiao Tong Univ, Shanghai Peoples Hosp 9, Dept Infect Dis, Sch Med, Shanghai 200011, Peoples R China

[6] Chinese Acad Sci, Inst Plant Physiol & Ecol, Shanghai Inst Biol Sci, Key Lab Synthet Biol, Shanghai 200032, Peoples R China

[7] Chinese Natl Human Genome Ctr Shanghai, Shanghai MOST Key Lab Hlth & Dis Genom, Shanghai 201203, Peoples R China

[8] Shanghai Key Lab Emergency Prevent Diag & Treatme, Shanghai, Peoples R China

来源：

JOURNAL OF INFECTION | 2021年 / 83卷 / 01期

基金：

中国国家自然科学基金;

关键词：

Tuberculosis; CRISPR-Cas12b; Mycobacterium tuberculosis; IS6110; Diagnosis; TB-QUICK; NUCLEIC-ACID DETECTION; PULMONARY TUBERCULOSIS; XPERT MTB/RIF; DNA; ENDONUCLEASE; PREVALENCE; CULTURE; PATIENT; BLOOD; ASSAY;

D O I：

10.1016/j.jinf.2021.04.032

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

Objectives: Tuberculosis (TB) remains one of the public health problems worldwide. Rapid, sensitive and cost-effective diagnosis of Mycobacterium tuberculosis (M.tb) is critical for TB control. Methods: We developed a novel M.tb DNA detection platform (nominated as TB-QUICK) which combined loop-mediated isothermal amplification (LAMP) and CRISPR-Cas12b detection. TB-QUICK was performed on pulmonary or plasma samples collected from 138 pulmonary TB (PTB) patients, 21 non-TB patients and 61 close contacts to TB patients. Acid-fast bacillus (AFB) smear, M.tb culture and GeneXpert MTB/RIF (Xpert) assays were routinely conducted in parallel. Results: By targeting M.tb IS6110, TB-QUICK platform could detect as low as 1.3 copy/mu L M.tb DNA within 2 h. In pulmonary TB samples, TB-QUICK exhibited improved overall sensitivity of 86.8% over M.tb culture (66.7%) and Xpert (70.4%), with the specificity of 95.2%. More significantly, TB-QUICK exhibited a superior sensitivity in AFB-negative samples (80.5%) compared to Xpert (57.1%) and M.tb culture (46.2%). In the detection of plasma M.tb DNA by TB-QUICK, 41.2% sensitivity for AFB-positive and 31.7% for AFB-negative patients were achieved. Conclusion: In conclusion, TB-QUICK exhibits rapidity and sensitivity for M.tb DNA detection with the superiority in smear-negative paucibacillary TB patients. The clinical application of TB-QUICK in TB diagnosis needs to be further validated in larger cohort. (C) 2021 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

引用

页码：54 / 60

页数：7

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