Volume 1, Issue 2 (4-2011)                   NCMBJ 2011, 1(2): 55-60 | Back to browse issues page

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Zaker Bostanabad S, Jabbarzadeh E, Pourazar S, Ghalami M. Typing of Mycobacterium tuberculosis Isolated from Patients in the Tehran City by Spoilgotyping. NCMBJ. 2011; 1 (2) :55-60
URL: http://ncmbjpiau.ir/article-1-23-en.html
Islamic Azad University Parand branch, Science Faculty, Biology Department , Jabbarzadeh@yahoo.com
Abstract:   (28416 Views)
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Aim and Background. spoilgotyping is a method based on 36bp DR(Direct Repeat) chromosomal loci polymorphism which is connected to one or two 35- 41 spacersequences. There have been 94 differet intra DR spacer sequences identified and only 43 of them are used as usual. Mycobacterium tuberculosis complex strains can be identified in order to lacking or having these sequences.

 

Materials and methods. spoilgotyping test was done on 149 TB smear positive patients.Primary separation of mycobacterium strains was done through petrof 4% method and by Lowenstein Jensen(LJ) media. Biochemical tests such as Niacin test / Catalase activity / Nitrate reduction were done due to strains identifying. Drug sensitivity to INH(2Mg/ml) / RIF(40Mg/ml) / STM(10Mg/ml) and ETBI(2Mg/ml) was accomplished proportional and strains were distributed to three groups: sensitive, MDR, non MDR. DNA was extracted by CTAB method on positive clonies. Sequences were amplified by PCR and after denaturating amplified sequences, hybridation with Streptavidine peroxidase enzyme through line reverse blot method was accomplished. Luminoscense was added and the membrane was put onto the X-ray film. Then we did radiology step.

 

Results: Serotypes were divided into 9 groups (Beijine/CAS23/Haarlem/U/T2/T1/EAI3/EAI2/CAS2). The majority group was Haarlem (27%) and minority one was T2 (0.4%). There were also identified 2 strains belonging to Mycobacterium bovis.

 

Conclusion: Used method in this study is easy/precipitate and has high sensitivity. In every time using this method there could be 43 samples distinguished.

 
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Type of Study: Research Article | Subject: Genetics
Received: 2011/05/5 | Accepted: 2014/04/23 | Published: 2014/04/23

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