PMID- 37821806
OWN - NLM
STAT- MEDLINE
DCOM- 20231101
LR  - 20231120
IS  - 1471-2334 (Electronic)
IS  - 1471-2334 (Linking)
VI  - 23
IP  - 1
DP  - 2023 Oct 11
TI  - A new method for the detection of Mycobacterium tuberculosis based on the 
      CRISPR/Cas system.
PG  - 680
LID - 10.1186/s12879-023-08656-4 [doi]
LID - 680
AB  - OBJECT: Mycobacterium tuberculosis (MTB) is a bacterium that can cause zoonoses 
      by aerosol transmission. Tuberculosis (TB) caused by MTB heavily burdens world 
      public health security. Developing efficient, specific, convenient, and 
      inexpensive MTB assays are essential for preventing and controlling TB. METHODS: 
      In this study, we established a specific detection method for MTB using the 
      Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR) system, 
      combined with recombinase mediated isothermal nucleic acid amplification (RAA) to 
      improve the sensitivity of the detection system and achieve "two-level" 
      amplification of the detection signal. The sensitivity and specificity of RAA 
      combined with the CRISPR/Cas system were analyzed. Using BACTEC 960 culture as 
      the gold standard for detecting MTB, we established the TB-CRISPR technique by 
      testing 504 samples from patients with suspected TB. RESULTS: MTB H37Ra could be 
      seen as low as 3.13 CFU/mL by the CRISPR-Cas12a system targeting IS6110. With 
      BACTEC960 culture (120 positives and 384 negatives) as the gold standard, the 
      sensitivity of the TB-CRISPR technique was 0.883 (0.809-0.932), and the 
      specificity was 0.940 (0.910-0.961). According to the receiver operating 
      characteristic (ROC) curve analysis, the area under the curve (AUC) reached 0.944 
      (0.914-0.975) within 95% CI. The positive likelihood ratio (PLR) was 14.747 
      (9.870-22.035), and the negative likelihood ratio (NLR) was 0.124 (0.076-0.203). 
      The positive predictive value (PPV) was 0.822 (0.742-0.881), and the negative 
      predictive value (NPV) was 0.963 (0.937-0.979). CONCLUSION: TB-CRISPR plays an 
      essential role in the molecular diagnosis of TB. The whole detection time is less 
      than 1.5 h. It is easy to operate and does not need complex instruments. It is of 
      great significance for the rapid detection of MTB and the clinical diagnosis of 
      TB.
CI  - (c) 2023. BioMed Central Ltd., part of Springer Nature.
FAU - Zhang, Xiaoyu
AU  - Zhang X
AD  - Department of Medical Laboratory, Weifang Medical University, Weifang, 261041, 
      Shandong, China.
FAU - He, Xiaoying
AU  - He X
AD  - Department of Laboratory Medicine, the Second People's Hospital of Weifang 
      261041, Weifang, 261041, Shandong, China.
FAU - Zhang, Yubo
AU  - Zhang Y
AD  - Department of Medical Laboratory, Weifang Medical University, Weifang, 261041, 
      Shandong, China.
FAU - Chen, Lei
AU  - Chen L
AD  - Department of Laboratory Medicine, the Second People's Hospital of Weifang 
      261041, Weifang, 261041, Shandong, China.
FAU - Pan, Zhaobao
AU  - Pan Z
AD  - Department of Laboratory Medicine, the Second People's Hospital of Weifang 
      261041, Weifang, 261041, Shandong, China.
FAU - Huang, Yueying
AU  - Huang Y
AD  - Department of Laboratory Medicine, Peking University Shenzhen Hospital, Shenzhen, 
      China.
FAU - Li, Heng
AU  - Li H
AD  - Department of Medical Laboratory, Weifang Medical University, Weifang, 261041, 
      Shandong, China. Leehome1985@163.com.
LA  - eng
PT  - Journal Article
DEP - 20231011
PL  - England
TA  - BMC Infect Dis
JT  - BMC infectious diseases
JID - 100968551
SB  - IM
MH  - Humans
MH  - *Mycobacterium tuberculosis/genetics
MH  - *Tuberculosis, Pulmonary/diagnosis
MH  - CRISPR-Cas Systems
MH  - *Tuberculosis/diagnosis/microbiology
MH  - Sensitivity and Specificity
PMC - PMC10568934
OTO - NOTNLM
OT  - CRISPR
OT  - Diagnosis
OT  - Molecular testing
OT  - Mycobacterium tuberculosis
OT  - RAA
COIS- The authors declare no conflict of interest. The funders had no role in the 
      study's design; in the collection, analyses, or interpretation of data; in the 
      writing of the manuscript, or in the decision to publish the results. The authors 
      declare no competing interests.
EDAT- 2023/10/12 00:42
MHDA- 2023/11/01 12:44
PMCR- 2023/10/11
CRDT- 2023/10/11 23:45
PHST- 2023/02/15 00:00 [received]
PHST- 2023/09/28 00:00 [accepted]
PHST- 2023/11/01 12:44 [medline]
PHST- 2023/10/12 00:42 [pubmed]
PHST- 2023/10/11 23:45 [entrez]
PHST- 2023/10/11 00:00 [pmc-release]
AID - 10.1186/s12879-023-08656-4 [pii]
AID - 8656 [pii]
AID - 10.1186/s12879-023-08656-4 [doi]
PST - epublish
SO  - BMC Infect Dis. 2023 Oct 11;23(1):680. doi: 10.1186/s12879-023-08656-4.