Cargando…

Rapid Detection of vanA Resistance Gene from E. faecalis Clinical Isolates Using Duplex Loop-Mediated Isothermal Amplification and Triplex PCR Assay

Today, the spread of vancomycin-resistant strains isolated from Enterococcus faecalis (E. faecalis) has become a major health concern worldwide. Therefore, it is essential to provide a rapid and sensitive assay for identifying vanA gene for timely and appropriate antimicrobial control of resistant e...

Descripción completa

Detalles Bibliográficos
Autores principales: Azizi, Mohsen, Motamedi, Hamid, Hossainpour, Hadi, Abiri, Ramin, Kashef, Mahsa, Ahmadi, Kamal, Moradi, Jale, Alvandi, Amirhooshang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9514927/
https://www.ncbi.nlm.nih.gov/pubmed/36177055
http://dx.doi.org/10.1155/2022/4384196
_version_ 1784798378184409088
author Azizi, Mohsen
Motamedi, Hamid
Hossainpour, Hadi
Abiri, Ramin
Kashef, Mahsa
Ahmadi, Kamal
Moradi, Jale
Alvandi, Amirhooshang
author_facet Azizi, Mohsen
Motamedi, Hamid
Hossainpour, Hadi
Abiri, Ramin
Kashef, Mahsa
Ahmadi, Kamal
Moradi, Jale
Alvandi, Amirhooshang
author_sort Azizi, Mohsen
collection PubMed
description Today, the spread of vancomycin-resistant strains isolated from Enterococcus faecalis (E. faecalis) has become a major health concern worldwide. Therefore, it is essential to provide a rapid and sensitive assay for identifying vanA gene for timely and appropriate antimicrobial control of resistant enterococcal infections. For this purpose, a cross-sectional study was performed on different clinical specimens of enterococci from Imam Reza hospital, Kermanshah, Iran. The antimicrobial susceptibility testing was determined by disk diffusion and MIC methods. Triplex-PCR and duplex-LAMP assays were also used to identify vanA E. faecalis resistance gene isolates. The results of this study shown that out of 108 Enterococcus isolates, 86, 18, 2, 1, and one isolates of E. faecalis, E. faecium, E. avium, E. psudoavium, and E. raffinosus were identified, respectively. On the other hand, E. faecalis was confirmed in 87 and 88 isolates using duplex-LAMP and triplex PCR, respectively. The LAMP primer set designed in this study can reliably identify seven distinct regions of the vanA gene, and finally the sensitivity, specificity, and the positive and negative predictive values of LAMP assay were shown to be 94.19%, 72.73%, 76.19%, and 93.10%, respectively. In general, sample processing, isothermal reaction and result reporting were completed using the LAMP assay in 75 minutes. Our findings suggest that LAMP assay has been approved as an alternative to the vancomycin resistance Enterococcus genotype (vanA and vanB) compared to other methods and has the advantage of being rapid, time-consuming, and easy for diagnosis.
format Online
Article
Text
id pubmed-9514927
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Hindawi
record_format MEDLINE/PubMed
spelling pubmed-95149272022-09-28 Rapid Detection of vanA Resistance Gene from E. faecalis Clinical Isolates Using Duplex Loop-Mediated Isothermal Amplification and Triplex PCR Assay Azizi, Mohsen Motamedi, Hamid Hossainpour, Hadi Abiri, Ramin Kashef, Mahsa Ahmadi, Kamal Moradi, Jale Alvandi, Amirhooshang Biomed Res Int Research Article Today, the spread of vancomycin-resistant strains isolated from Enterococcus faecalis (E. faecalis) has become a major health concern worldwide. Therefore, it is essential to provide a rapid and sensitive assay for identifying vanA gene for timely and appropriate antimicrobial control of resistant enterococcal infections. For this purpose, a cross-sectional study was performed on different clinical specimens of enterococci from Imam Reza hospital, Kermanshah, Iran. The antimicrobial susceptibility testing was determined by disk diffusion and MIC methods. Triplex-PCR and duplex-LAMP assays were also used to identify vanA E. faecalis resistance gene isolates. The results of this study shown that out of 108 Enterococcus isolates, 86, 18, 2, 1, and one isolates of E. faecalis, E. faecium, E. avium, E. psudoavium, and E. raffinosus were identified, respectively. On the other hand, E. faecalis was confirmed in 87 and 88 isolates using duplex-LAMP and triplex PCR, respectively. The LAMP primer set designed in this study can reliably identify seven distinct regions of the vanA gene, and finally the sensitivity, specificity, and the positive and negative predictive values of LAMP assay were shown to be 94.19%, 72.73%, 76.19%, and 93.10%, respectively. In general, sample processing, isothermal reaction and result reporting were completed using the LAMP assay in 75 minutes. Our findings suggest that LAMP assay has been approved as an alternative to the vancomycin resistance Enterococcus genotype (vanA and vanB) compared to other methods and has the advantage of being rapid, time-consuming, and easy for diagnosis. Hindawi 2022-09-20 /pmc/articles/PMC9514927/ /pubmed/36177055 http://dx.doi.org/10.1155/2022/4384196 Text en Copyright © 2022 Mohsen Azizi et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Azizi, Mohsen
Motamedi, Hamid
Hossainpour, Hadi
Abiri, Ramin
Kashef, Mahsa
Ahmadi, Kamal
Moradi, Jale
Alvandi, Amirhooshang
Rapid Detection of vanA Resistance Gene from E. faecalis Clinical Isolates Using Duplex Loop-Mediated Isothermal Amplification and Triplex PCR Assay
title Rapid Detection of vanA Resistance Gene from E. faecalis Clinical Isolates Using Duplex Loop-Mediated Isothermal Amplification and Triplex PCR Assay
title_full Rapid Detection of vanA Resistance Gene from E. faecalis Clinical Isolates Using Duplex Loop-Mediated Isothermal Amplification and Triplex PCR Assay
title_fullStr Rapid Detection of vanA Resistance Gene from E. faecalis Clinical Isolates Using Duplex Loop-Mediated Isothermal Amplification and Triplex PCR Assay
title_full_unstemmed Rapid Detection of vanA Resistance Gene from E. faecalis Clinical Isolates Using Duplex Loop-Mediated Isothermal Amplification and Triplex PCR Assay
title_short Rapid Detection of vanA Resistance Gene from E. faecalis Clinical Isolates Using Duplex Loop-Mediated Isothermal Amplification and Triplex PCR Assay
title_sort rapid detection of vana resistance gene from e. faecalis clinical isolates using duplex loop-mediated isothermal amplification and triplex pcr assay
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9514927/
https://www.ncbi.nlm.nih.gov/pubmed/36177055
http://dx.doi.org/10.1155/2022/4384196
work_keys_str_mv AT azizimohsen rapiddetectionofvanaresistancegenefromefaecalisclinicalisolatesusingduplexloopmediatedisothermalamplificationandtriplexpcrassay
AT motamedihamid rapiddetectionofvanaresistancegenefromefaecalisclinicalisolatesusingduplexloopmediatedisothermalamplificationandtriplexpcrassay
AT hossainpourhadi rapiddetectionofvanaresistancegenefromefaecalisclinicalisolatesusingduplexloopmediatedisothermalamplificationandtriplexpcrassay
AT abiriramin rapiddetectionofvanaresistancegenefromefaecalisclinicalisolatesusingduplexloopmediatedisothermalamplificationandtriplexpcrassay
AT kashefmahsa rapiddetectionofvanaresistancegenefromefaecalisclinicalisolatesusingduplexloopmediatedisothermalamplificationandtriplexpcrassay
AT ahmadikamal rapiddetectionofvanaresistancegenefromefaecalisclinicalisolatesusingduplexloopmediatedisothermalamplificationandtriplexpcrassay
AT moradijale rapiddetectionofvanaresistancegenefromefaecalisclinicalisolatesusingduplexloopmediatedisothermalamplificationandtriplexpcrassay
AT alvandiamirhooshang rapiddetectionofvanaresistancegenefromefaecalisclinicalisolatesusingduplexloopmediatedisothermalamplificationandtriplexpcrassay