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. 2016 Summer;11(3):238-247.

Species-specific PCR for the Diagnosis and Determination of Antibiotic Susceptibilities of Brucella Strains Isolated from Tehran, Iran

Gholam Reza Irajian  1 , Faramarz Masjedian Jazi  1 , Reza Mirnejad  2 , Vahhab Piranfar  2 , Taghi Zahraei Salehi  3 , Noor Amir Mozafari  1 , Ehsanollah Ghaznavi-Rad  4 , Mahmoud Khormali  3
Affiliations

Species-specific PCR for the Diagnosis and Determination of Antibiotic Susceptibilities of Brucella Strains Isolated from Tehran, Iran

Gholam Reza Irajian et al. Iran J Pathol. 2016 Summer.

Abstract

Background: Brucellosis is an endemic zoonotic disease in the Middle East. This study intended to design a uniplex PCR assay for the detection and differentiation of Brucella at the species level and determining the antibiotic susceptibility pattern of Brucella in Iran.

Methods: Sixty-eight Brucella specimens (38 animal and 30 human specimens) were analyzed using PCR (using one pair of primers). Antibiotic susceptibility patterns were evaluated and compared using the E-Test and disk diffusion susceptibility test. Tigecycline susceptibility pattern was compared with other antibiotics.

Results: Thirty six isolates of B. melitensis, 2 isolates of B. abortus and 1 isolate of B. suis from the 38 animal specimens, 24 isolates of B. melitensis and 6 isolates of B. abortus from the 30 human specimens were differentiated. The MIC50 values of doxycycline for human and animal specimens were 125 and 10 μg/ml, respectively, tigecycline 0.064 μg/ml for human specimens and 0.125μg/ml for animal specimens, and trimethoprim/ sulfamethoxazole and ciprofloxacin 0.065 and 0.125μg/ml, respectively, for both human and animal specimens. The highest MIC50 value of streptomycin in the human specimens was 0.5μg/ml and 1μg/ml for the animal specimens. The greatest resistance shown was to tetracycline and gentamicin, respectively.

Conclusion: Uniplex PCR for the detection and differentiation of Brucella at the strain level is faster and less expensive than multiplex PCR, and the antibiotics doxycycline, rifampin, trimethoprim-sulfamethoxazole, ciprofloxacin, and ofloxacin are the most effective antibiotics for treating brucellosis. Resistance to tigecycline is increasing, and we recommend that it be used in a combination regimen.

Keywords: Antibiotic Susceptibilities; Brucella; Tigecycline; Uniplex PCR.

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Figures

Fig. 1
Fig. 1
Agarose gel electrophoresis of B. melitensis, B. abortus and B.suis strains. (M: DNA molecular marker, bp: base pair). Lane 1: animal B. suis, 770 bp, Lane 2: B. melitasis 16M 398 bp, Lane 3: animal B. melitensis and Lane 9: Hu- man B. melitasis. Lane 4: B. abortus S19, 520 bp. Lane 5: animal B. abortus. Lane 6-8: human B. abortus, negative control not shown in Fig. 1
See this image and copyright information in PMC

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