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

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...

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Autores principales: Moeini-Zanjani, Ali, Pournajaf, Abazar, Ferdosi-Shahandashti, Elaheh, Gholami, Mehrdad, Masjedian, Faramarz, Khafri, Soraya, Rajabnia, Ramazan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7666441/
https://www.ncbi.nlm.nih.gov/pubmed/33187548
http://dx.doi.org/10.1186/s13104-020-05377-8
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author Moeini-Zanjani, Ali
Pournajaf, Abazar
Ferdosi-Shahandashti, Elaheh
Gholami, Mehrdad
Masjedian, Faramarz
Khafri, Soraya
Rajabnia, Ramazan
author_facet Moeini-Zanjani, Ali
Pournajaf, Abazar
Ferdosi-Shahandashti, Elaheh
Gholami, Mehrdad
Masjedian, Faramarz
Khafri, Soraya
Rajabnia, Ramazan
author_sort Moeini-Zanjani, Ali
collection PubMed
description 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.
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spelling pubmed-76664412020-11-16 Comparison of loop-mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species Moeini-Zanjani, Ali Pournajaf, Abazar Ferdosi-Shahandashti, Elaheh Gholami, Mehrdad Masjedian, Faramarz Khafri, Soraya Rajabnia, Ramazan BMC Res Notes Research Note 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. BioMed Central 2020-11-13 /pmc/articles/PMC7666441/ /pubmed/33187548 http://dx.doi.org/10.1186/s13104-020-05377-8 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Note
Moeini-Zanjani, Ali
Pournajaf, Abazar
Ferdosi-Shahandashti, Elaheh
Gholami, Mehrdad
Masjedian, Faramarz
Khafri, Soraya
Rajabnia, Ramazan
Comparison of loop-mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species
title Comparison of loop-mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species
title_full Comparison of loop-mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species
title_fullStr Comparison of loop-mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species
title_full_unstemmed Comparison of loop-mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species
title_short Comparison of loop-mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species
title_sort comparison of loop-mediated isothermal amplification and conventional pcr tests for diagnosis of common brucella species
topic Research Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7666441/
https://www.ncbi.nlm.nih.gov/pubmed/33187548
http://dx.doi.org/10.1186/s13104-020-05377-8
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