An Isothermal Method for Sensitive Detection of Mycobacterium tuberculosis Complex Using Clustered Regularly Interspaced Short Palindromic Repeats/Cas12a Cis and Trans Cleavage - Details

author：

Indexed by：

Scopus

Abstract：

Tuberculosis　is　one　of　the　most　serious　infectious　diseases,　resulting　in　death　worldwide.　Traditional　detection　methods　are　not　enough　to　meet　the　clinical　requirements　of　rapid　diagnosis,　high　specificity,　and　high　sensitivity.　Fast,　sensitive,　and　accurate　detection　of　Mycobacterium　tuberculosis　(MTB)　is　urgently　needed　to　treat　and　control　tuberculosis　disease.　Clustered　regularly　interspaced　short　palindromic　repeats　(CRISPR)–associated　proteins　(Cas12a)　exhibit　strong　nonspecific　degradation　ability　of　exogenous　single-strand　nucleic　acids　(trans　cleavage)　after　specific　recognition　of　target　sequence.　We　purified　Cas12a　protein　and　selected　a　proper　guide　RNA　based　on　conserved　sequences　of　MTB　from　designed　guide　RNA　library.　Then,　we　proposed　a　novel　detection　method　based　on　recombinase　polymerase　amplification　and　CRISPR/Cas12a　nuclease　system　for　specific　and　sensitive　detection　of　MTB　DNA.　The　assay,　based　on　fluorescence　detection,　showed　4.48　fmol/L　of　limit　of　detection　and　good　linear　correlation　of　concentration　with　fluorescence　value　(R2　=　0.9775).　It　also　showed　good　performance　in　distinguishing　other　bacteria.　Furthermore,　its　clinical　performance　was　evaluated　by　193　samples　and　showed　sensitivity　of　99.29%　(139/140)　and　specificity　of　100%　(53/53)　at　99%　CI,　compared　with　culture　method.　Taken　together,　the　CRISPR/Cas12a　system　showed　good　specificity,　excellent　sensitivity,　and　excellent　accuracy　for　MTB　detection,　and　it　meets　requirements　of　MTB　detection　in　clinical　samples　and　has　great　potential　for　clinical　translation.　©　2020　Association　for　Molecular　Pathology　and　American　Society　for　Investigative　Pathology

Keyword：

Community：

[ 1 ] [Xu, H.]The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, China
[ 2 ] [Xu, H.]Mengchao Med-X Center, Fuzhou University, Fuzhou, China
[ 3 ] [Zhang, X.]The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, China
[ 4 ] [Zhang, X.]Mengchao Med-X Center, Fuzhou University, Fuzhou, China
[ 5 ] [Cai, Z.]The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, China
[ 6 ] [Cai, Z.]Mengchao Med-X Center, Fuzhou University, Fuzhou, China
[ 7 ] [Cai, Z.]School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, China
[ 8 ] [Dong, X.]The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, China
[ 9 ] [Dong, X.]Mengchao Med-X Center, Fuzhou University, Fuzhou, China
[ 10 ] [Chen, G.]The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, China
[ 11 ] [Chen, G.]Mengchao Med-X Center, Fuzhou University, Fuzhou, China
[ 12 ] [Chen, G.]School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, China
[ 13 ] [Li, Z.]The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, China
[ 14 ] [Li, Z.]Mengchao Med-X Center, Fuzhou University, Fuzhou, China
[ 15 ] [Qiu, L.]The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, China
[ 16 ] [Qiu, L.]Mengchao Med-X Center, Fuzhou University, Fuzhou, China
[ 17 ] [He, L.]College of Biological Science and Engineering, Fuzhou University, Fuzhou, China
[ 18 ] [Liang, B.]Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, China
[ 19 ] [Liu, X.]The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, China
[ 20 ] [Liu, X.]Mengchao Med-X Center, Fuzhou University, Fuzhou, China
[ 21 ] [Liu, X.]School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, China
[ 22 ] [Liu, J.]The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, China
[ 23 ] [Liu, J.]Mengchao Med-X Center, Fuzhou University, Fuzhou, China
[ 24 ] [Liu, J.]Liver Disease Center, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China

Reprint 's Address：

[Liu, X.]Mengchao Hepatobiliary Hospital of Fujian Medical UniversityChina

Email：

liuxl@fjirsm.ac.cn

Show more details

Related Keywords：

Source ：

Journal of Molecular Diagnostics

ISSN： 1525-1578

Year： 2020

Issue： 8

Volume： 22

Page： 1020-1029

5 . 5 6 8

JCR@2020

3 . 4 0 0

JCR@2023

ESI HC Threshold：142

JCR Journal Grade：1

CAS Journal Grade：2

Cited Count：

WoS CC Cited Count：

SCOPUS Cited Count：

ESI Highly Cited Papers on the List： 0 Unfold All

WanFang Cited Count：

Chinese Cited Count：

30 Days PV： 1

Affiliated Colleges：

Get Fulltext

DOI Library Discovery Baidu Scholar Search SCOPUS

Type
Departments

All Years Choose Year From to