Comparison of loop-mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species
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RESEARCH NOTE
Comparison of loop‑mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species Ali Moeini‑Zanjani1, Abazar Pournajaf1,2, Elaheh Ferdosi‑Shahandashti2,3, Mehrdad Gholami4, Faramarz Masjedian5, Soraya Khafri6 and Ramazan Rajabnia1,2*
Abstract Objective: Rapid, reliable, and affordable detection of Brucella species via the molecular methods remains a chal‑ lenge. In recent years, loop-mediated isothermal amplification (LAMP) is a functional nucleic acid amplification technique offering a substitute to polymerase chain reaction (PCR). So, we compared the LAMP assay with the con‑ ventional PCR for the identification of common Brucella species in Iran. In this study, LAMP assay was comprehensively evaluated against the common PCR method. A group of specific LAMP primers were used to amplify a highly specific fragment from the sequence of the Brucella abortus, bcsp31 gene. Sensitivity and specificity values of tests were done with a set of 78 (50 Brucella and 28 non-Brucella) strains. Results: A dilution series of B. abortus DNA indicated that the LAMP reaction could reliably detect 10 (fg/µl) DNA target copies per reaction within 36 min, which is 10 times greater than the PCR assay. In summary, we conclude that LAMP assay provide accurate and fast test results to identify of common Brucella species in low-complexity labs, mainly in low and lower middle income countries. Keyword: Brucellosis, Diagnosis, PCR, LAMP Introduction Brucella species are small, coccobacilli, Gram-negative, absolute aerobic and non-moving bacteria which causes undulant fever in humans and leads to abortion and infertility in animals [1]. Brucella can be transmitted to humans through direct contact with animals or their products that are contaminated with these bacteria [2]. Brucella genus has six species that cannot be distinguished from each other due to the close phenotypic and antigenic similarities with conventional microbiological methods [3, 4]. From these six species, [B. abortus, B. melitensis, B. suis and B. canis] generally causes human *Correspondence: [email protected] 2 Infectious Diseases and Tropical Medicine Research Center, Babol University of Medical Sciences, Babol, Iran Full list of author information is available at the end of the article
infection [5]. About half a million cases of human brucellosis are reported annually which is estimated to be 10–25% less than the real number of the 1997 world health organization (WHO) report [6]. The centers for disease control and prevention strategic planning group as listed B. abortus, B. melitensis, B. suis as category B biothreat agents [5, 7]. There are currently three main methods for the identification of brucellosis [8]. Overall, in human brucellosis, isolation of the bacterium by blood culture is considered as the gold standard for laboratory diagnosis, but the procedure is time consuming [9, 10], and has a low sensitivity in the range of 15–70% depending on the bacterial species and infe
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