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:: Volume 20, Issue 1 (Scientific Journal of Kurdistan University of Medical Sciences 2015) ::
SJKU 2015, 20(1): 10-17 Back to browse issues page
Simultaneous identification and comparison of pathogenic Brucella species in human serum and blood samples by means of multiplex PCR and confirmation of the results by PCR-RFLP
Elnaz Akbarzadeh 1, Jamileh Noroosi Dr1, Vahab Piranfar 2, Dariush Ghasemi 2, Shiva Mirkalantari Dr3, Reza Mirnejad Dr 4
1- Islamic Azad University, Tehran North Branch
2- Baqiyatallah University of Medical Sciences
3- Semnan University of Medical Sciences
4- Baqiyatallah University of Medical Sciences , rmirnejad@yahoo.com
Abstract:   (3944 Views)
Background and Aim: There are few studies on the isolation of Brucella abortus and Brucella melitensis from patients´ sera. The aim of this study was simultaneous isolation of pathogenic brucella species from human serum samples by multiplex PCR method. Material andMethods: 50 blood and serum samples isolated from patients with clinically suspected brucellosis were inoculated into brucella agar medium and we used multiplex-PCR, with three primers to detect brucella species. We incubated 0.5 ml of serum in Brucella broth for 72 hours at 37 °C in 5% carbon dioxide. To confirm the results of PCR, the PCR products were restricted by restriction enzymes, TaqI and RsaI. Result: From 50 blood samples 4 (8%) cultures were positive. Using biochemical tests and after determination of the characteristics of the positive cases, they were identified as B. melitensis. After multiplex PCR, 9 cases (18%) were positive and 41 cases (82%) were negative. Among the positive sera, B. melitensis was identified in 7 cases (78%) and B. abortus in 2 cases (22%). Of 50 blood samples, 5 (10%) were positive and 45 (90%) were negative for B. melitensis. All the results were confirmed by PCR-RFLP. Conclusion: Our study showed that multiplex PCR method was simple, rapid and is more sensitive for isolation of brucella from serum (especially B. abortus and B. melitensis) in comparison to complete blood. Keywords: Brucella spp., Multiplex-PCR, Serum blood, PCR-RFLP Received: Jul 7, 2014 Accepted: Dec 10, 2014
Keywords: Brucella spp., Multiplex-PCR, Serum blood, PCR-RFLP
Full-Text [PDF 1257 kb]   (1095 Downloads)    
Type of Study: Research | Subject: General
Received: 2015/03/14 | Accepted: 2015/03/14 | Published: 2015/03/14
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Volume 20, Issue 1 (Scientific Journal of Kurdistan University of Medical Sciences 2015) Back to browse issues page
مجله علمی دانشگاه علوم پزشکی کردستان Scientific Journal of Kurdistan University of Medical Sciences
مجله علمی دانشگاه علوم پزشکی کردستان Scientific Journal of Kurdistan University of Medical Sciences
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