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

被引:18
|
作者
Moeini-Zanjani, Ali [1 ]
Pournajaf, Abazar [1 ,2 ]
Ferdosi-Shahandashti, Elaheh [2 ,3 ]
Gholami, Mehrdad [4 ]
Masjedian, Faramarz [5 ]
Khafri, Soraya [6 ]
Rajabnia, Ramazan [1 ,2 ]
机构
[1] Babol Univ Med Sci, Fac Med, Dept Microbiol, Babol, Iran
[2] Babol Univ Med Sci, Infect Dis & Trop Med Res Ctr, Babol, Iran
[3] Babol Univ Med Sci, Fac Med, Dept Med Biotechnol, Babol, Iran
[4] Mazandaran Univ Med Sci, Fac Med, Dept Microbiol & Virol, Sari, Iran
[5] Iran Univ Med Sci, Fac Med, Dept Microbiol, Tehran, Iran
[6] Babol Univ Med Sci, Fac Med, Dept Biostat & Epidemiol, Babol, Iran
关键词
Brucellosis; Diagnosis; PCR; LAMP; RAPID DETECTION; MELITENSIS;
D O I
10.1186/s13104-020-05377-8
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
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/mu l) 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.
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页数:5
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