Species Identification Of Mycobacterium And Preliminary Study On The Proteome Of Mycobacterium Tuberculosis Beijing Family

Objective To investigate the species of non-tuberculosis mycobac-terium (NTM) and subspecies of Mycobacterium chelonae/abscessus group preliminarily and to study the molecular methods for species identification of mycobacterium. To preliminary analyze the proteome of Mycobacterium tuberculosis Beijing family and Mycobacterium bovis Bacille Calmette-Guerin (BCG) strain and their different protein expression.Methods Verified identification of iaolates identified as NTM by local laboratories were performed by means of L-J, PNB/TCH solid culture in our own laboratory. Then, species of NTM were identified by hsp65 and rpoB PCR-RFLP methods. Subspecies of M. chelonae/ abscessus group were identified by bacteriological characters and muti-line probe reversed hybridization method and muti-targets, including 16s rRNA,16s-23s rRNA, rpoB, hsp65, recA and sod A genes, sequencing and PCR-RFLP analysis. The proteome were studied by two-dimensional gel electrophoresis (TDE) and Matrix-assisted laser desorption-Ionization time of flight (MALDI-TOF) methods.Results 178 clinical isolates were identified to be 27 M. tuberculosis strains and 1 M. bovis strain and 150 NTM strains by our verified tests. 150 NTM strains identified were including 72 Mycobacterium intracell-ural,21 Mycobacterium chelonae/abscessus group,12 Mycobacterium gordonae,11 Mycobacterium avium,9 Mycobacterium fortuitum,2 Mycobacterium kansasii,1 Mycobacterium simiae,1 Mycobacterium arupense,1 Mycobacterium brumae and 14 iaolates not identified to species level.27 Mycobacterium chelonae/abscessus group strains were identified to three species, including 19 M. abscessus subspecies M. abscessus,3 M. abscessus subspecies M. massilense and 5 Mycobacterium fukienense as a novel species closely related to M. chelonae-M. abscessus group by further identification. By compared analysis proteome,765 proteins of M. tuberculosis Beijing family strain CCDC5079 were analysed.6 proteins were uniquely expressed and 2 proteins were highly expressed in CCDC5079 strain compared with BCG strain.Conclusions Lots of species of NTM could course chronic lung disease in China, and Mycobacterium intracellural and Mycobacterium abscessus group are the main prevalent Mycobacterium pathogenic bacterias. Muti-targets sequcing method may identify NTM to species or subspecies effectively. There are at least three subspecies closely related to M. chelonae/abscessus group to course chronic lung disease in China. We described Mycobacterium fukienense as a novel species firstly in the word. There was a significant difference between proteomes of M. tuberculosis Beijing family strain and BCG strain.