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
Author(s):
Abstract:
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
Language:
Persian
Published:
Scientific Journal of Kurdistan University of Medical Sciences, Volume:20 Issue: 1, 2015
Pages:
10 to 17
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