The Efficacy of Multiplex PCR in Comparison with Agglutination and ELISA in Diagnosis of Human Brucellosis

Reza  Shahrokhabadi; Ebrahim Rahimi; Hassan Mommtaz; Rahele Poursahebi; Somayeh Doostmohamadi

Zahedan Journal of Research in Medical Sciences

The Official Journal of Zahedan University of Medical Sciences

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The Efficacy of Multiplex PCR in Comparison with Agglutination and ELISA in Diagnosis of Human Brucellosis

Author(s):

Reza Shahrokhabadi^1,*,

Ebrahim Rahimi²,

Hassan Mommtaz³,

Rahele Poursahebi⁴,

Somayeh Doostmohamadi⁵

1Young Researchers Club, Veterinary Medicine, Islamic Azad University, Shahr-e-Kord Branch, Shahr-e-Kord, Iran

2Department of Food Hygiene, Faculty of Veterinary M edicine Shahr-e-Kord Branch, Islamic Azad University, Shahr-e-Kord, Iran

3Department of Microbiology, Faculty of Veterinary M edicine Shahr-e-Kord Branch, Islamic Azad University, Shahr-e-Kord, Iran

4Department of Veterinary, Shahid Bahonar University, Kerman, Iran

5Department of Animal Sciences, Payam-e-Noor Univers tiy, Tehran, Iran

Zahedan Journal of Research in Medical Sciences:Vol. 16, issue 4; 16-28

Published online:May 05, 2013

Article type:Research Article

Received:Jan 04, 2013

Accepted:Feb 13, 2013

How to Cite:Reza ShahrokhabadiEbrahim RahimiHassan MommtazRahele PoursahebiSomayeh DoostmohamadiThe Efficacy of Multiplex PCR in Comparison with Agglutination and ELISA in Diagnosis of Human Brucellosis.Zahedan J Res Med Sci.16(4):16-28.

Abstract

Background: Human brucellosis is an endemic disease in many countries including Iran. Exact diagnosis of brucellosis is not just based on clinical symptoms, because it will be considered in differential diagnosis of other diseases. Therefore, defining organism in culture or identification of organism by serological and molecular methods for confirming clinical diagnosis is necessary. Our aim was to develop a diagnostic PCR assay and define the optimal clinical specimen for this test.

Materials and Methods: This cross-sectional and descriptive study was from February 2011 to November 2012. Results of standard agglutination test (SAT) and specific immunoglobulin IgG and IgM by enzyme-linked immunosorbent assay (ELISA) were compared with multiplex PCR in 116 patients with suspected brucellosis referred to the Ali Ebn-e-Abitaleb hospital, Rafsanjan, Iran. Their sera were collected and tested by SAT, ELISA and multiplex PCR. DNA was extracted from serum samples and examined by multiplex PCR involving specific primers for Brucella melitensis and Brucella abortus based on IS711 in the brucella chromosome.

Results: Brucellosis was confirmed in 116 patients (75% male and 25% female) based on applied diagnostic methods and clinical features. Results of ELISA, the SAT, and PCR were positive in 116, respectively. B. abortus and B. melitensis were detected in 101 and 15 patients.

Conclusion: The results of present study showed that multiplex PCR assay is a rapid and sensitive technique for diagnosis of brucellosis compared to SAT. However it is more accurate when coupled with conventional methods.

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