FLASH-PCR as a Simple and Efficient Method for Detection of <i>Brucella</i> spp. Infection

Abstract

Background: Brucella spp. are Gram-positive, rod-shaped, and spore-forming bacilli. Brucella abortus and B. melitensis are the main causes of brucellosis. Objectives: The aim of the study was to establish a rapid and simple molecular method for the detection of this disease. Methods: Forty-five Brucella spp. were isolated from blood samples using the BACTEC Fluorescent 9050 system and were detected by anti-IgM or IgG Brucella specific antigen. DNA extraction was conducted on all samples. Fluorescent amplification based specific hybridization (FLASH-PCR) test was utilized to detect the 351-bp fragment from eryD gene, which was specific for Brucella spp. Results: A 351-bp fragment resulted from PCR reaction and showed the accuracy of designed primers. This fragment was successfully amplified in the FLASH-PCR reaction. In this study, we have positive and negative samples and a standard method. In addition, we calculated the sensitivity and specificity of this method as 100%. Conclusions: Results of the study was proved that the FLASH-PCR method was a rapid, sensitive, and safe method for the detection of Brucella genome in whole blood samples of patient harbored brucellosis and is recommended for routine usage.