Comparison of loop-mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species.

Autor: Moeini-Zanjani A; Department of Microbiology, Faculty of Medicine, Babol University of Medical Sciences, Babol, Iran., Pournajaf A; Department of Microbiology, Faculty of Medicine, Babol University of Medical Sciences, Babol, Iran.; Infectious Diseases and Tropical Medicine Research Center, Babol University of Medical Sciences, Babol, Iran., Ferdosi-Shahandashti E; Infectious Diseases and Tropical Medicine Research Center, Babol University of Medical Sciences, Babol, Iran.; Department of Medical Biotechnology, Faculty of Medicine, Babol University of Medical Sciences, Babol, Iran., Gholami M; Department of Microbiology and Virology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran., Masjedian F; Department of Microbiology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran., Khafri S; Department of Biostatistics and Epidemiology, Faculty of Medicine, Babol University of Medical Sciences, Babol, Iran., Rajabnia R; Department of Microbiology, Faculty of Medicine, Babol University of Medical Sciences, Babol, Iran. ramazan69@yahoo.com.; Infectious Diseases and Tropical Medicine Research Center, Babol University of Medical Sciences, Babol, Iran. ramazan69@yahoo.com.
Jazyk: angličtina
Zdroj: BMC research notes [BMC Res Notes] 2020 Nov 13; Vol. 13 (1), pp. 533. Date of Electronic Publication: 2020 Nov 13.
DOI: 10.1186/s13104-020-05377-8
Abstrakt: Objective: Rapid, reliable, and affordable detection of Brucella species via the molecular methods remains a challenge. 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 conventional 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.
Databáze: MEDLINE
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