application of deletion- targeted multiplex pcr technique for detection of mycobacterium tuberculosis beijing strains in samples from tuberculosis patients.

نویسندگان

azar dokht khosravi health research institute, infectious and tropical diseases research center, ahvaz jundishapur university of medical sciences, ahvaz, iran and department of microbiology, school of medicine, ahvaz jundishapur university of medical sciences, ahvaz, iran.

hamed goodarzi department of microbiology, school of medicine, ahvaz jundishapur university of medical sciences, ahvaz, iran.

seyedmohammad alavi health research institute, infectious and tropical diseases research center, ahvaz jundishapur university of medical sciences, ahvaz, iran.

mohammadreza akhond department of statistics, mathematical science and computer faculty, shahid chamran university, ahvaz, iran.

چکیده

background and objective: molecular epidemiological studies have shown that certain genotypes of mycobacterium tuberculosis (mtb) are over-represented in limited geographical regions, suggesting of evolution of certain genotypes with increasing virulence and pathogenicity. beijing strain of mtb was initially described by its potential to cause outbreaks worldwide and its association with drug resistance. due to tuberculosis (tb)-related mortality which is associated with beijing genotype , this study was designed with the aim to detect the mtb beijing genotype in the region of study. materials and methods: a total of 170 clinical isolates of mtb were collected from the tb reference laboratory of khuzestan province, iran, over one year period from february 2010 to february 2011. phenotypic tests were used for preliminary detection of mtb. culture positive mtb isolates were confirmed by multiplex pcr based on is 6110 gene with subsequent  screening  for resistance to isoniazid (inh), and rifampin (rif) by pcr using relevant primers. three set of primers were used to differentiate beijing from non-beijing strains by using deletion- targeted multiplex  (dtm) pcr. results: from 160 pcr-confirmed mtb isolates, 18 (11.25%) showed mutation in kat g gene related to inh resistance and 20 (12.5%), associated with mutation in rpo b gene related to rif resistance, and 8 (5%) were detected as beijing strain using multiplex pcr. the majority of detected beijing strains (6/8[75%]) comprised mutation in kat g gene with the prevalent mutation specifically in codon 315. in 4 beijing strains (2.5%), mutation in rpo b gene were also detected. conclusion: using dtm- pcr, the rate of beijing strains in the region of study was determined as 5%. although for detection of mtb antimicrobial resistance, it is advised to use a combination of conventional antimicrobial susceptibility testing and molecular techniques, however for time saving, it seems that dtm-pcr, is a simple technique for use in areas of the world where beijing strains are highly prevalent.

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عنوان ژورنال:
iranian journal of microbiology

جلد ۶، شماره ۵، صفحات ۳۳۰-۳۳۴

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