Real-Time Loop-Mediated Isothermal Amplification (LAMP) Method for Quantitative Salmonella Typhi Detection Based on ViaB Gene
Background & Objective: The Salmonella origin diseases is wide which could be from gastroenteritis, enteric fever, bacteraemia and focal infection. The infection of salmonella usually comes from the consumption of the contaminated food or water with animal or human excrement. This study aimed to...
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| Format: | Article |
| Language: | English |
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Fasa University of Medical Sciences
2021-05-01
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| Series: | Journal of Advanced Biomedical Sciences |
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| Online Access: | http://jabs.fums.ac.ir/article-1-2546-en.pdf |
| _version_ | 1797679542783967232 |
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| author | Mahdi Sohrab Keivan Majidzadeh Abbas Morovvati Mohammad Soleimani |
| author_facet | Mahdi Sohrab Keivan Majidzadeh Abbas Morovvati Mohammad Soleimani |
| author_sort | Mahdi Sohrab |
| collection | DOAJ |
| description | Background & Objective: The Salmonella origin diseases is wide which could be from gastroenteritis, enteric fever, bacteraemia and focal infection. The infection of salmonella usually comes from the consumption of the contaminated food or water with animal or human excrement. This study aimed to design a Quantitative-LAMP method to real-time detection of Salmonella enterica serovar Typhi (S. Typhi), the causative agent of Typhoid fever.
Materials & Methods: A new LAMP primer set specifically was designed based on ViaB gene and used for detection of S. Typhi. To evaluate the analytical specificity, the genome of some Salmonella-related and non-related bacteria were subjected to the S. Typhi LAMP assay. Also, the analytical sensitivity or limit of detection of the assay was evaluated. Furthermore, in the experiment, turbidity in tubes was assessed by a turbidimeter and then a standard curve was depicted by plotting time threshold values against the log of ViaB gene copy number. With this standard curve, we could use the method to make a quantitative method
Results: The Salmonella Typhi LAMP assay specifically assessed the ViaB gene. The analytical sensitivity of the assay when using agarose gel electrophoresis or amplification plot obtained from Loop amp real-time turbidimeter system was 0.28 fg whereas with direct observation of fluorescent color change to evaluate amplification was 2.8 fg. So, the lower limit of detection of the assay when applying the revealing methods was ~1 and 8 copies of the ViaB gene respectively.
Conclusions: The Salmonella Typhi LAMP assay is a simple and accurate tool to detect the causative agent of Typhoid fever that may designate to apply in clinical laboratories |
| first_indexed | 2024-03-11T23:16:18Z |
| format | Article |
| id | doaj.art-267370a4e3654bc5be24bf47d3766255 |
| institution | Directory Open Access Journal |
| issn | 2228-5105 2783-1523 |
| language | English |
| last_indexed | 2024-03-11T23:16:18Z |
| publishDate | 2021-05-01 |
| publisher | Fasa University of Medical Sciences |
| record_format | Article |
| series | Journal of Advanced Biomedical Sciences |
| spelling | doaj.art-267370a4e3654bc5be24bf47d37662552023-09-20T21:28:33ZengFasa University of Medical SciencesJournal of Advanced Biomedical Sciences2228-51052783-15232021-05-0111238313838Real-Time Loop-Mediated Isothermal Amplification (LAMP) Method for Quantitative Salmonella Typhi Detection Based on ViaB GeneMahdi Sohrab0Keivan Majidzadeh1Abbas Morovvati2Mohammad Soleimani3 Department of Microbiology, Islamic Azad University, Qom Branch, Qom, Iran Breast Cancer Research Center (BCRC), Academic Center for Education, Culture & Research (ACECR), Tehran, Iran Department of Microbiology, Islamic Azad University, Qom Branch, Qom, Iran Department of Microbiology, Faculty of Medicine, AJA University of Medical Sciences, Tehran, Iran Background & Objective: The Salmonella origin diseases is wide which could be from gastroenteritis, enteric fever, bacteraemia and focal infection. The infection of salmonella usually comes from the consumption of the contaminated food or water with animal or human excrement. This study aimed to design a Quantitative-LAMP method to real-time detection of Salmonella enterica serovar Typhi (S. Typhi), the causative agent of Typhoid fever. Materials & Methods: A new LAMP primer set specifically was designed based on ViaB gene and used for detection of S. Typhi. To evaluate the analytical specificity, the genome of some Salmonella-related and non-related bacteria were subjected to the S. Typhi LAMP assay. Also, the analytical sensitivity or limit of detection of the assay was evaluated. Furthermore, in the experiment, turbidity in tubes was assessed by a turbidimeter and then a standard curve was depicted by plotting time threshold values against the log of ViaB gene copy number. With this standard curve, we could use the method to make a quantitative method Results: The Salmonella Typhi LAMP assay specifically assessed the ViaB gene. The analytical sensitivity of the assay when using agarose gel electrophoresis or amplification plot obtained from Loop amp real-time turbidimeter system was 0.28 fg whereas with direct observation of fluorescent color change to evaluate amplification was 2.8 fg. So, the lower limit of detection of the assay when applying the revealing methods was ~1 and 8 copies of the ViaB gene respectively. Conclusions: The Salmonella Typhi LAMP assay is a simple and accurate tool to detect the causative agent of Typhoid fever that may designate to apply in clinical laboratorieshttp://jabs.fums.ac.ir/article-1-2546-en.pdflampsalmonella typhityphoid feverspecificity and sensitivity |
| spellingShingle | Mahdi Sohrab Keivan Majidzadeh Abbas Morovvati Mohammad Soleimani Real-Time Loop-Mediated Isothermal Amplification (LAMP) Method for Quantitative Salmonella Typhi Detection Based on ViaB Gene Journal of Advanced Biomedical Sciences lamp salmonella typhi typhoid fever specificity and sensitivity |
| title | Real-Time Loop-Mediated Isothermal Amplification (LAMP) Method for Quantitative Salmonella Typhi Detection Based on ViaB Gene |
| title_full | Real-Time Loop-Mediated Isothermal Amplification (LAMP) Method for Quantitative Salmonella Typhi Detection Based on ViaB Gene |
| title_fullStr | Real-Time Loop-Mediated Isothermal Amplification (LAMP) Method for Quantitative Salmonella Typhi Detection Based on ViaB Gene |
| title_full_unstemmed | Real-Time Loop-Mediated Isothermal Amplification (LAMP) Method for Quantitative Salmonella Typhi Detection Based on ViaB Gene |
| title_short | Real-Time Loop-Mediated Isothermal Amplification (LAMP) Method for Quantitative Salmonella Typhi Detection Based on ViaB Gene |
| title_sort | real time loop mediated isothermal amplification lamp method for quantitative salmonella typhi detection based on viab gene |
| topic | lamp salmonella typhi typhoid fever specificity and sensitivity |
| url | http://jabs.fums.ac.ir/article-1-2546-en.pdf |
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