Identification of Nontuberculous Mycobacteria in Fish Tank Water by Phenotyping and PCR-RFLP and DNA sequencing of Mycobacterial hsp65 gene

Abstract

Abstract: Objective To investigate NTM distribution in fish tank water in Beijing area. Methods A total of 30 fish tank water samples were collected and incubated for NTM growth in Lowenstein-Jensen media at 28℃ and 37℃,respectively.NTM were further identified by three methods:(1)traditional phenotypic methods-growth and biochemical characteristics;(2)restriction fragment length polymorphism(RFLP) of a PCR-amplified 439-bp segment of the gene encoding the 65-kDa heat shock protein(hsp65);(3) PCR-sequencing. Results Thirty-five NTM strains were isolated in 29 out of 30 fish tank water samples.Traditional phenotypic Methods and genotypic characterization(RFLP and sequencing) revealed that 8 strains(23%) were M.Gordonae,8 strains(23%) belonged to M.fortuitum complex(2 strains M.fortuitum and 2 strains of M.peregrinum),9 strains(26%) were M.genavense, 1 strain was M.nonchromogenicum,and 9 strains were unidentified NTM. Conclusion NTM were wildly distributed in fish tank water that might be one of major sources for NTM infection.PCR-based molecular Methods showed high sensitivity and specificity in identification of NTMs.

Key words: Nontuberculous mycobacteria, PCR, Restriction fragment length polymorphism

Chen Xue¹,Cai Lin,Zhao Ting,et al.. Identification of Nontuberculous Mycobacteria in Fish Tank Water by Phenotyping and PCR-RFLP and DNA sequencing of Mycobacterial hsp65 gene[J]. Chinese Journal of Antituberculosis, 2007, 29(2): 154-159.

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