title: [Identification and evaluation of a new nucleic acid amplification test target for specific detection of Mycobacterium tuberculosis complex].
abstract: OBJECTIVE
  To identify and evaluate a new nucleic acid amplification (NAA) test target for specific detection of Mycobacterium tuberculosis (MTB) complex (MTC).
  
  
  METHODS
  MTC-specific fragment was obtained by ISSR genotyping technology. Primer pairs were designed based on the sequences of MTC-specific fragment and tested in 211 mycobacterial strains including 107 MTC strains and 104 nontuberculous mycobacteria (NTM) strains. IS6110 element (specific identification of MTC strains) and 16s rRNA gene (specific identification of Mycobacterium) amplification were used as a control to evaluate the efficacy of the NAA test target in the detection of MTC strains.
  
  
  RESULTS
  One MTC-specific fragment with the length of 588 bp, located in 315947 - 316534 of the genome from MTB reference strain H(37) Rv, were obtained, cloned and sequenced. MTC-specific primer pairs MTCF/R were designed based on these sequences. All 211 mycobacterial strains accurately produced the genus-specific 16s rRNA amplicon. All MTC strains were positive in the MTCF/R PCR amplification while 99% MTC strains (106/107) were positive in the amplification of IS6110 sequences. All NTM strains were negative in both IS6110 and MTCF/R PCR amplification.
  
  
  CONCLUSIONS
  The MTC-specific fragment developed in this study can be used as a new NAA test target to correctly distinguish MTC from NTM.
authors: Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai 200433, China.
authors: Shi-hui
authors: SH
authors: Gao
authors: Ying-wei
authors: YW
authors: Zhao
authors: Zhong-yi
authors: ZY
authors: Hu
authors: Jie
authors: J
authors: Wang
authors: Jun-mei
authors: JM
authors: Lu
authors: Li-ping
authors: LP
authors: Yan
authors: Qi
authors: Q
authors: Guo
authors: Hui
authors: H
authors: Ma
authors: Lian-hua
authors: LH
authors: Qin
mesh_terms: D001483:Base Sequence; D004269:DNA, Bacterial; D005838:Genotype; D009161:Mycobacterium / Q000145:classification / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques
chemical_list: D004269:DNA, Bacterial