Differentiation of Mycobacterium tuberculosis clinical strains using restrikction fragment length polymorphism technique based on hsp65 gene.
Journal Title: مجله دانشگاه علوم پزشکی شهرکرد - Year 2005, Vol 7, Issue 3
Abstract
Background and aim: Polymerase chain reaction (PCR) coupled with Restriction Fragment Length Polymorphism analysis (RFLP) has been used for genotyping of Mycobacteria in recent years. While detailed differentiation of non tuberculosis Mycobacteria has been described by this technique, even in some species ability of this technique for molecular epidemiology has been proven, only a small number of Mycobacterium tuberculosis (MTB) strains have been studied. The present study was conducted to apply this method to an expanded population of MTB isolates. Methods: A total of 150 clinical isolates were collected from patients referred to TB reference unit, PHLS, Ahvaz, Iran. Acid fast staining was performed for the isolates and they were identified as MTB by using conventional culture and biochemical tests. The PCR-RFLP method uses a simple DNA extraction followed by a PCR step based on amplification of a 439 bp fragment of hsp65 gene involving genus specific primers. The restriction enzyme analysis was then performed by digestion of products with HaeIII and BstEII enzymes. Results: Our results showed that 145 clinical isolates (96.6%) showed the identical restriction patterns similar to M. tuberculosis reference strains equal to 165/140/72 bp fragments for HaeIII and 250/120/82 bp fragments for BstEII digests. Three different patterns were observed for five clinical strains in HaeIII digest as 165/145 bp (three isolates), 180/100/80 bp (one isolate) and 194/72 bp (one isolate), while their BstEII digestion patterns showed no variation and were similar to other isolates. Conclusion: Our results showed that in clinical strains of mycobacterium tuberculosis, few polymorphisms in hsp65 fragment might be present.
Authors and Affiliations
azardokht Khosravi, abdorrazagh Hashemi
Keywords
Related Articles
- EP ID EP82445
- DOI -
- Views 98
- Downloads 0
Evaluation of WHO’s partogram alert line for prediction of the APGAR score at the first minute after birth
Background and aim: The early diagnosis of abnormal progress of labour and prevention of long term delivery are the main keys to reduce fetal mortality rate due to hypoxia and tissue ischemia. One of the WHO’s proposed m...
The comparison of the effects of both propranolol and transcutaneous electrical nerve stimulation in prevention of migraine headache
Background and aim: Migraine headache is a common familial disorder and its prevalence varies base on sex, age and races. There are two types of treatments for migraine: acute headache treatment (abortive) and preventive...
بررسی اثر تزريق موضعي دگزامتازون و بوپي واكائين بر شدت درد پس از عمل تانسیلکتومی
زمينه و هدف: تانسيلكتومي از شايع ترين اعمال جراحي است كه داراي عوارضي همانند درد، خونريزي، لارنژيت و لارنگواسپاسم مي باشد. كنترل درد بدليل كاهش رنج و بيقراري در كودك و كاهش احتمال خونريزي بعد از عمل از اهميت خاصي برخوردا...
The hepatotoxicity of aqueous methanol extract and fractionated-methanol extract of Phytolacca americana in male rat using perfusion method
Background and aim: Phytolacca americana is a native plant in several states of USA and mostly its boiled leaves are used as a popular salad (called grandmother salad) in American diet. It grows widely in the coastal are...
Effects of aspirin on teratogenic characteristic of alcohol in limb morphoyenesis of mouse embryo
Abnormalities caused by alcohol are very extensive and are called fetal alcoholic syndrome (FAS). There are some hypotheses about pathological mechanisms of alcohol. Based on one of the recent hypotheses, alcohol consump...