PMID- 35508977
OWN - NLM
STAT- MEDLINE
DCOM- 20220506
LR  - 20220716
IS  - 1471-2334 (Electronic)
IS  - 1471-2334 (Linking)
VI  - 22
IP  - 1
DP  - 2022 May 4
TI  - CRISPR-Cas12a combination to alleviate the false-positive in loop-mediated 
      isothermal amplification-based diagnosis of Neisseria meningitidis.
PG  - 429
LID - 10.1186/s12879-022-07363-w [doi]
LID - 429
AB  - BACKGROUND: Loop isothermal amplification (LAMP) has recently been proposed as a 
      point-of-care diagnostic tool to detect acute infectious pathogens; however, this 
      technique embeds risk of generating false-positive results. Whereas, with 
      abilities to accurately recognize specific sequence, the CRISPR/Cas12a can forms 
      complexes with cognate RNA sensors and cleave pathogen's DNA targets 
      complimerntary to its cognate RNA, afterward acquiring the collateral activity to 
      unbiasedly cut nearby off-target fragments. Therefore, if relevant 
      fluorescent-quencher-nucleic probes are present in the reaction, the non-specific 
      cleavage of probes releases fluorescences and establish diagnostic read-outs. 
      METHODS: The MetA gene of N. meningitidis was selected as target to optimize the 
      LAMP reaction, whereas pseudo-dilution series of N. meningitidis gemonics DNA was 
      used to establish the detection limit of LAMP/Cas12a combination assay. The 
      diagnostic performance of established LAMP/Cas12a combination assay was validated 
      in comparation with standard real-time PCR on 51 CSF samples (14 N. meningitidis 
      confirmed patients and 37 control subjects). RESULTS: In relevant biochemical 
      conditions, CRISPR-Cas12a and LAMP can work synchronously to accurately identify 
      genetics materials of Nesseria menitigistis at the level 40 copies/reaction less 
      than 2 h. CONCLUSIONS: In properly optimized conditions, the CRISPR-Cas12a system 
      helps to alleviate false positive result hence enhancing the specificity of the 
      LAMP assays.
CI  - (c) 2022. The Author(s).
FAU - Trung, Ngo Tat
AU  - Trung NT
AD  - Centre for Genetics Consultation and Cancer Screening (CGC), Hanoi, Vietnam. 
      tatrungngo@gmail.com.
AD  - Vietnamese-German Center for Medical Research (VG-CARE), Hanoi, Vietnam. 
      tatrungngo@gmail.com.
AD  - Faculty of Tropical and Infectious Diseases, 108 Military Central Hospital, Hai 
      Ba Trung District, No 1, Tran Hung Dao Street, Hanoi, Vietnam. 
      tatrungngo@gmail.com.
AD  - 108 Institute of Clinical Medical and Pharmaceutical Sciences, Hanoi, Vietnam. 
      tatrungngo@gmail.com.
FAU - Son, Le Huu Phuc
AU  - Son LHP
AD  - Centre for Genetics Consultation and Cancer Screening (CGC), Hanoi, Vietnam.
AD  - Vietnamese-German Center for Medical Research (VG-CARE), Hanoi, Vietnam.
FAU - Hien, Trinh Xuan
AU  - Hien TX
AD  - Centre for Genetics Consultation and Cancer Screening (CGC), Hanoi, Vietnam.
AD  - Vietnamese-German Center for Medical Research (VG-CARE), Hanoi, Vietnam.
FAU - Quyen, Dao Thanh
AU  - Quyen DT
AD  - Vietnamese-German Center for Medical Research (VG-CARE), Hanoi, Vietnam.
AD  - Faculty of Tropical and Infectious Diseases, 108 Military Central Hospital, Hai 
      Ba Trung District, No 1, Tran Hung Dao Street, Hanoi, Vietnam.
AD  - 108 Institute of Clinical Medical and Pharmaceutical Sciences, Hanoi, Vietnam.
FAU - Bang, Mai Hong
AU  - Bang MH
AD  - Vietnamese-German Center for Medical Research (VG-CARE), Hanoi, Vietnam.
AD  - 108 Institute of Clinical Medical and Pharmaceutical Sciences, Hanoi, Vietnam.
FAU - Song, Le Huu
AU  - Song LH
AD  - Vietnamese-German Center for Medical Research (VG-CARE), Hanoi, Vietnam. 
      lehuusong@108-icid.com.
AD  - Faculty of Tropical and Infectious Diseases, 108 Military Central Hospital, Hai 
      Ba Trung District, No 1, Tran Hung Dao Street, Hanoi, Vietnam. 
      lehuusong@108-icid.com.
AD  - 108 Institute of Clinical Medical and Pharmaceutical Sciences, Hanoi, Vietnam. 
      lehuusong@108-icid.com.
LA  - eng
GR  - 108.06-2017.21/Vietnam National Foundation for Science and Technology Development 
      (NAFOSTED)/
GR  - 364/2020/HD-NCKHCN/The Vietnamese Ministry of National Defence/
PT  - Journal Article
DEP - 20220504
PL  - England
TA  - BMC Infect Dis
JT  - BMC infectious diseases
JID - 100968551
RN  - 63231-63-0 (RNA)
RN  - 9007-49-2 (DNA)
RN  - LAMP assay
SB  - IM
MH  - *CRISPR-Cas Systems
MH  - DNA
MH  - Humans
MH  - Molecular Diagnostic Techniques
MH  - *Neisseria meningitidis/genetics
MH  - Nucleic Acid Amplification Techniques/methods
MH  - RNA
PMC - PMC9066958
OTO - NOTNLM
OT  - CRISPR
OT  - Cas12a
OT  - LAMP
OT  - Nesseria menitigistis
OT  - PCR
COIS- The authors declare no conflict of interests.
EDAT- 2022/05/06 06:00
MHDA- 2022/05/07 06:00
PMCR- 2022/05/04
CRDT- 2022/05/05 00:03
PHST- 2020/11/17 00:00 [received]
PHST- 2022/04/07 00:00 [accepted]
PHST- 2022/05/05 00:03 [entrez]
PHST- 2022/05/06 06:00 [pubmed]
PHST- 2022/05/07 06:00 [medline]
PHST- 2022/05/04 00:00 [pmc-release]
AID - 10.1186/s12879-022-07363-w [pii]
AID - 7363 [pii]
AID - 10.1186/s12879-022-07363-w [doi]
PST - epublish
SO  - BMC Infect Dis. 2022 May 4;22(1):429. doi: 10.1186/s12879-022-07363-w.