Typing of Mycobacterium tuberculosis Isolated fromPatients in the Tehran City by Spoilgotyping

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 ifferet intraDR 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 LowensteinJensen(LJ) media. Biochemical tests such as Niacin test/Catalase activity/Nitrate reduction were done dueto 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 afterdenaturating amplified sequences, hybridation with Streptavidine peroxidase enzyme through line reverseblot 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). Themajority group was Haarlem (27%) and minority one was T2(0.4%). There were also identified 2 strainsbelonging to Mycobacterium bovis.Conclusion. Used method in this study is easy/precipitate and has high sensitivity. In every time usindthis method there could be 43 samples distinguished.