PMID- 29875194
OWN - NLM
STAT- MEDLINE
DCOM- 20190418
LR  - 20220408
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Print)
IS  - 0095-1137 (Linking)
VI  - 56
IP  - 8
DP  - 2018 Aug
TI  - McSpoligotyping, a One-Step Melting Curve Analysis-Based Protocol for 
      Spoligotyping of Mycobacterium tuberculosis.
LID - 10.1128/JCM.00539-18 [doi]
LID - e00539-18
AB  - The direct repeat (DR) region in the Mycobacterium tuberculosis (MTB) genome is 
      composed of highly polymorphic direct variant repeats, which are the basis of 
      spacer oligonucleotide typing (spoligotyping) to study the population structure 
      and epidemiology of M. tuberculosis However, the membrane hybridization-based 
      detection format requires various post-PCR manipulations and is prone to 
      carryover contamination, restricting its wide use in high-TB-burden and 
      resource-limited countries. We developed a one-step spoligotyping protocol, 
      termed McSpoligotyping, based on real-time PCR. The typing results can be 
      generated within 3 h by a single step of DNA addition. When evaluated with a 
      collection of 1,968 isolates of MTB, McSpoligotyping agreed 97.71% (1,923/1,968) 
      by sample and 99.93% (84,568/84,624) by spacer with traditional spoligotyping. 
      Sequencing results showed that McSpoligotyping was even more accurate than 
      spoligotyping (99.34% versus 98.37%). Further exploration of the false results of 
      McSpoligotyping revealed the presence of single-nucleotide polymorphisms in the 
      DR region. We concluded that McSpoligotyping could be used in epidemiology 
      studies of tuberculosis by taking advantage of the shortened procedure, ease of 
      use, and compatibility of results with standard spoligotyping.
CI  - Copyright (c) 2018 American Society for Microbiology.
FAU - Zeng, Xiaohong
AU  - Zeng X
AD  - State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School 
      of Public Health, Xiamen University, Xiamen, Fujian, China.
AD  - Engineering Research Centre of Molecular Diagnostics, Ministry of Education, 
      State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen 
      University, Xiamen, Fujian, China.
AD  - Engineering Research Centre of Personalized Molecular Diagnostics of Xiamen, 
      Fujian, China.
FAU - Xu, Ye
AU  - Xu Y
AD  - State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School 
      of Public Health, Xiamen University, Xiamen, Fujian, China.
AD  - Engineering Research Centre of Molecular Diagnostics, Ministry of Education, 
      State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen 
      University, Xiamen, Fujian, China.
AD  - Engineering Research Centre of Personalized Molecular Diagnostics of Xiamen, 
      Fujian, China.
FAU - Zhou, Yang
AU  - Zhou Y
AD  - National Tuberculosis Reference Laboratory, National Center for Tuberculosis 
      Control and Prevention, Chinese Center for Disease Control and Prevention, 
      Beijing, China.
FAU - Li, Hui
AU  - Li H
AD  - Tuberculosis Reference Laboratory, Henan Center for Disease Control and 
      Prevention, Zhengzhou, Henan, China.
FAU - Zheng, Rongrong
AU  - Zheng R
AD  - Division of Tuberculosis Control and Prevention, Xiamen Center for Disease 
      Control and Prevention, Xiamen, Fujian, China.
FAU - Tan, Yaoju
AU  - Tan Y
AD  - Department of Clinical Laboratory, Guangzhou Chest Hospital, Guangzhou, China.
FAU - Sun, Bingqi
AU  - Sun B
AD  - Tuberculosis Laboratory, Shenyang Chest Hospital, Shenyang, Liaoning, China.
FAU - Zhao, Yanlin
AU  - Zhao Y
AD  - National Tuberculosis Reference Laboratory, National Center for Tuberculosis 
      Control and Prevention, Chinese Center for Disease Control and Prevention, 
      Beijing, China zhaoyl@chinacdc.cn qgli@xmu.edu.cn.
FAU - Li, Qingge
AU  - Li Q
AD  - State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School 
      of Public Health, Xiamen University, Xiamen, Fujian, China zhaoyl@chinacdc.cn 
      qgli@xmu.edu.cn.
AD  - Engineering Research Centre of Molecular Diagnostics, Ministry of Education, 
      State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen 
      University, Xiamen, Fujian, China.
AD  - Engineering Research Centre of Personalized Molecular Diagnostics of Xiamen, 
      Fujian, China.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20180726
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 0 (DNA, Bacterial)
SB  - IM
MH  - DNA, Bacterial/genetics
MH  - Genotype
MH  - Humans
MH  - Molecular Epidemiology/*methods
MH  - Molecular Typing/*methods/standards
MH  - Mycobacterium tuberculosis/*classification/genetics
MH  - Polymorphism, Single Nucleotide
MH  - Real-Time Polymerase Chain Reaction
MH  - Repetitive Sequences, Nucleic Acid/genetics
MH  - Sequence Analysis, DNA
MH  - Time Factors
MH  - Tuberculosis/*microbiology
PMC - PMC6062818
OTO - NOTNLM
OT  - Mycobacterium tuberculosis
OT  - automatic interpretation
OT  - melting curve analysis
OT  - one step
OT  - spoligotyping
EDAT- 2018/06/08 06:00
MHDA- 2019/04/19 06:00
PMCR- 2019/01/26
CRDT- 2018/06/08 06:00
PHST- 2018/03/31 00:00 [received]
PHST- 2018/05/31 00:00 [accepted]
PHST- 2018/06/08 06:00 [pubmed]
PHST- 2019/04/19 06:00 [medline]
PHST- 2018/06/08 06:00 [entrez]
PHST- 2019/01/26 00:00 [pmc-release]
AID - JCM.00539-18 [pii]
AID - 00539-18 [pii]
AID - 10.1128/JCM.00539-18 [doi]
PST - epublish
SO  - J Clin Microbiol. 2018 Jul 26;56(8):e00539-18. doi: 10.1128/JCM.00539-18. Print 
      2018 Aug.