Hashemi Abodolrazagh, Shojaei Hasan, Heidarieh Parvin, Aslani Mohamad Mehdi, Naser Abass Daei
Ahvaz Jundishapur University of Medical Sciences, Iran.
New Microbiol. 2012 Jan;35(1):61-5. Epub 2012 Jan 10.
In the current study we aimed to execute a rather less complicated molecular tying method, i.e. the random amplification of polymorphic DNA (RAPD) analysis to find the heterogeneity of Iranian strains of Mycobacterium tuberculosis. The isolates comprised a total of 96 strains of M. tuberculosis collected from clinical specimens of patients in Isfahan and Tehran. The isolates were assigned to the species M. tuberculosis by the key conventional and molecular methods. They were then subjected to RAPD analysis by four arbitrary primers, namely, the primers 27F, 1525R, MS- GF and INS-2. They were then evaluated for the number and intensity of the band patterns. The RAPD profiles of the Iranian isolates showed a degree of heterogeneity which varied based on the primer used. However, analysis of the isolates by primer INS-2 revealed the highest degree of diversity yielding 31 distinguishable RAPD types. RAPD analysis provides a rapid and easy means of identifying heterogeneity among the M. tuberculosis isolates. This typing system might be considered a valuable alternative molecular typing for countries with limited resources provided that the reproducibility and reliability of the method is carefully assured.
在本研究中,我们旨在实施一种相对不太复杂的分子分型方法,即随机扩增多态性DNA(RAPD)分析,以发现伊朗结核分枝杆菌菌株的异质性。分离株共有96株结核分枝杆菌,从伊斯法罕和德黑兰患者的临床标本中收集。通过关键的传统和分子方法将分离株鉴定为结核分枝杆菌。然后用四个任意引物,即引物27F、1525R、MS-GF和INS-2对它们进行RAPD分析。然后评估条带模式的数量和强度。伊朗分离株的RAPD图谱显示出一定程度的异质性,其因所用引物而异。然而,用引物INS-2对分离株进行分析时,发现多样性程度最高,产生了31种可区分的RAPD类型。RAPD分析为鉴定结核分枝杆菌分离株之间的异质性提供了一种快速简便的方法。只要仔细确保该方法的可重复性和可靠性,这种分型系统可能被认为是资源有限国家一种有价值的替代分子分型方法。