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:: Volume 11, Issue 4 (winter 2011) ::
J Ardabil Univ Med Sci 2011, 11(4): 345-353 Back to browse issues page
Detection of Amikacin-Resistance among MDR Strains of Mycobacterium tuberculosis by Using PCR-RFLP Method
Parisa Tahmasebi , Fatemeh Maryam Sheikolslami, Parisa Farnia, Majid Sadeghizadeh, Rashid Ramazanzadeh, Mahdi Kazempoor, MohammadReza Masjedi, Ali Akbar Velayati
Abstract:   (6413 Views)

  Background & objectives : Amikacin is one of the key second-line drugs for treatment of tuberculosis. Mutations at the codons 1400, 1401and1483 of the 16srRNA gene are associated with resistance to amikacin. The purpose of this study was to detect these mutations using PCR-RFLP method in multi-drug resistant (MDR) strains of Mycobacterium tuberculosis showing resistance to amikacin.

  Methods : Susceptibility of strains (n=100) against first and second–line anti-tuberculosis drugs was performed by proportional method. Based on antimicrobial resistance pattern 97 strains were analyzed by PCR-RFLP method. rrs1096 and rrs1539 primers were used to amplify a 460bp region of the rrs gene. Then, the PCR products were digested using Tai 1 and Dde1 restriction enzymes. The results were analyzed by the SPSS software using Chi-square test.

  Results : Based on results from proportional method, 63 strains (64.9%) were MDR (Multiple Drug Resistant), 26 (26.8%) and 8 (8.2%) strains were susceptible and non-MDR, respectively. Also, 13.4% and 6.1% of the strains were XDR (Extensively Drug Resistant) and TDR ( Totally drug resistant ) respectively. Using PCR-RFLP method, 7 (7.2%) strains were resistant and 90 (92.7%) strains were susceptible to amikacin respectively. Moreover, we found that the mutation at the codon 1400 was the most frequent mutations responsible for resistance to amikacin.

  Conclusion : The PCR-RFLP method can be used as a supplemental method to detect resistance to amikacin however to increase our knowledge, mutatuions in several number of codons in rrs gene need to be studied.

Keywords: PCR-RFLP; Mycobacterium tuberculosis; MDR; rrs; Amikacin
Full-Text [PDF 247 kb]   (2662 Downloads)    
Type of Study: article | Subject: Special
Received: 2011/08/1 | Accepted: 2011/11/7 | Published: 2011/12/22
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Tahmasebi P, Sheikolslami F M, Farnia P, Sadeghizadeh M, Ramazanzadeh R, Kazempoor M, et al . Detection of Amikacin-Resistance among MDR Strains of Mycobacterium tuberculosis by Using PCR-RFLP Method. J Ardabil Univ Med Sci. 2011; 11 (4) :345-353
URL: http://jarums.arums.ac.ir/article-1-163-en.html


Volume 11, Issue 4 (winter 2011) Back to browse issues page
مجله دانشگاه علوم پزشکی اردبیل Journal of Ardabil University of Medical Sciences
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