<sec><title>Objectives</title><p>Brucellosis is a major zoonotic disease that poses a significant public health threat worldwide. The classical bacteriological detection process used to identify <italic>Brucella</italic> spp. is difficult and time-consuming. This study aimed to develop a novel molecular assay for detecting brucellosis.</p></sec><sec><title>Methods</title><p>All complete sequences of chromosome 1 with 2.1-Mbp lengths were compared among all available <italic>Brucella</italic> sequences. A unique repeat sequence (URS) locus on chromosome 1 could differentiate <italic>Brucella abortus</italic> from <italic>Brucella melitensis</italic>. A primer set was designed to flank the unique locus. A total of 136 lymph nodes and blood samples were evaluated and classified by the URS-polymerase chain reaction (PCR) method in 2013–2014.</p></sec><sec><title>Results</title><p>Biochemical tests and bacteriophage typing as the golden standard indicated that all <italic>Brucella</italic> spp. isolates were <italic>B. melitensis</italic> biovar 1 and <italic>B. abortus</italic> biovar 3. The PCR results were the same as the bacteriological method for detecting <italic>Brucella</italic> spp. The sensitivity and specificity of the URS-PCR method make it suitable for detecting <italic>B. abortus</italic> and <italic>B. melitensis</italic>.</p></sec><sec><title>Conclusion</title><p>Quick detection of <italic>B. abortus</italic> and <italic>B. melitensis</italic> can provide the most effective strategies for control of these bacteria. The advantage of this method over other presented methods is that both <italic>B. abortus</italic> and <italic>B. melitensis</italic> are detectable in a single test tube. Furthermore, this method covered 100% of all <italic>B. melitensis</italic> and <italic>B. abortus</italic> biotypes. The development of this URS-PCR method is the first step toward the development of a novel kit for the molecular identification of <italic>B. abortus</italic> and <italic>B. melitensis</italic>.</p></sec>
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Objectives
Gastrointestinal tract infection is still one of the serious public health problems in many geographic areas and is endemic in most countries including Iran. Early detection of the gastrointestinal tract pathogens can be extremely important. The aim of the current study was to apply a shortened time-multiplex polymerase chain reaction (PCR) for rapid and simultaneous detection of <i>Salmonella</i> spp., <i>Shigella</i> spp., and <i>Vibrio cholera</i>. Methods
The standard and clinical strains of <i>Salmonella</i> spp., <i>Shigella</i> spp., and <i>V. cholerae</i> were used in the assay. Multiplex PCR was performed and optimized based on amplification of <i>invA</i>, putative integrase, and <i>ompW</i> genes for detecting <i>Salmonella</i> spp., <i>Shigella</i> spp., and <i>V. cholerae</i>, respectively. The specificity of the assay was evaluated by testing 12 different bacterial species. Results
Only <i>Salmonella</i> spp., <i>Shigella</i> spp., and <i>V. cholerae</i> strains had positive results when subjected to the assay using multiplex PCR. The assay showed a high sensitivity, and no amplification products were observed in multiplex PCR with any of the other microorganisms. Conclusion
Our study indicated that the <i>invA</i>, putative integrase, and <i>ompW</i>-based multiplex PCR assay appears to be an efficient method for rapid and simultaneous detection of <i>Salmonella</i> spp., <i>Shigella</i> spp., and <i>V. cholerae.</i>
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