Comparison of primers B4/B5 and F4/R2 for Detection of Brucella DNA in Serum by PCR

Introduction: Brucellosis is a worldwide zoonosis and is endemic in all parts of the Iran. The development of new diagnostic techniques that facilitate rapid detection and identification of brucellae and minimize the risk of laboratory infection is of great practical importance. The aim of this study was to evaluate two different PCR methods for the detection of Brucella spp. DNA in human serum samples. Methods: Serial dilutions of purified DNA were made. The two pairs of primers have chosen amplified regions of two different Brucella genes. All amplifications were performed in a total volume of 25 μl, containing serial dilutions of Brucella DNA and positive and negative controls. The negative control contained sterile water instead of DNA template. As positive controls, DNA isolated from Brucella spp was used. Results: The two PCR assays had excellent sensitivity for detection of Brucella spp DNA in serum samples. The two primers assayed did not show a difference in sensitivity for detecting purified Brucella DNA. They detecting purified DNA of Brucella in ranging between 4/25 ng to 4/25 fg. Conclusion: The sensitivity of two primers set did not show a difference in detecting of Brucella DNA in serum sample; however, as the primers B4/B5 amplified a shorter fragment in comparison to primers F4/R2, we think that they could provide a useful tool for diagnosis of human brucellosis