Detection of Shigella in Milk and Clinical Samples by Magnetic Immunocaptured-Loop-Mediated Isothermal Amplification Assay
Shigella is an important human food-borne zoonosis bacterial pathogen, and can cause clinically severe diarrhea. There is an urgent need to develop a specific, sensitive, and rapid methodology for detection of this pathogen. In this study, loop-mediated isothermal amplification (LAMP) combined with...
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doaj-a8f382930a864b5bae933edd8246e1f72020-11-24T22:38:35ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-02-01910.3389/fmicb.2018.00094313574Detection of Shigella in Milk and Clinical Samples by Magnetic Immunocaptured-Loop-Mediated Isothermal Amplification AssayLiding Zhang0Qiujiang Wei1Qinqin Han2Qiang Chen3Wenlin Tai4Jinyang Zhang5Yuzhu Song6Xueshan Xia7Molecular Medicine Research Center of Yunnan Province, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, ChinaKunming Biomed International, Kunming, ChinaMolecular Medicine Research Center of Yunnan Province, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, ChinaMolecular Medicine Research Center of Yunnan Province, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, ChinaYunnan Molecular Diagnosis Centre, The Second Affiliation Hospital of Kunming Medical University, Kunming, ChinaMolecular Medicine Research Center of Yunnan Province, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, ChinaMolecular Medicine Research Center of Yunnan Province, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, ChinaMolecular Medicine Research Center of Yunnan Province, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, ChinaShigella is an important human food-borne zoonosis bacterial pathogen, and can cause clinically severe diarrhea. There is an urgent need to develop a specific, sensitive, and rapid methodology for detection of this pathogen. In this study, loop-mediated isothermal amplification (LAMP) combined with magnetic immunocapture assay (IC-LAMP) was first developed for the detection of Shigella in pure culture, artificial milk, and clinical stool samples. This method exhibited a detection limit of 8.7 CFU/mL. Compared with polymerase chain reaction, IC-LAMP is sensitive, specific, and reliable for monitoring Shigella. Additionally, IC-LAMP is more convenient, efficient, and rapid than ordinary LAMP, as it is more efficiently enriches pathogen cells without extraction of genomic DNA. Under isothermal conditions, the amplification curves and the green fluorescence were detected within 30 min in the presence of genomic DNA template. The overall analysis time was approximately 1 h, including the enrichment and lysis of the bacterial cells, a significantly short detection time. Therefore, the IC-LAMP methodology described here is potentially useful for the efficient detection of Shigella in various samples.http://journal.frontiersin.org/article/10.3389/fmicb.2018.00094/fullShigellaIC-LAMPimmunocapturedetectionspecificity |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Liding Zhang Qiujiang Wei Qinqin Han Qiang Chen Wenlin Tai Jinyang Zhang Yuzhu Song Xueshan Xia |
spellingShingle |
Liding Zhang Qiujiang Wei Qinqin Han Qiang Chen Wenlin Tai Jinyang Zhang Yuzhu Song Xueshan Xia Detection of Shigella in Milk and Clinical Samples by Magnetic Immunocaptured-Loop-Mediated Isothermal Amplification Assay Frontiers in Microbiology Shigella IC-LAMP immunocapture detection specificity |
author_facet |
Liding Zhang Qiujiang Wei Qinqin Han Qiang Chen Wenlin Tai Jinyang Zhang Yuzhu Song Xueshan Xia |
author_sort |
Liding Zhang |
title |
Detection of Shigella in Milk and Clinical Samples by Magnetic Immunocaptured-Loop-Mediated Isothermal Amplification Assay |
title_short |
Detection of Shigella in Milk and Clinical Samples by Magnetic Immunocaptured-Loop-Mediated Isothermal Amplification Assay |
title_full |
Detection of Shigella in Milk and Clinical Samples by Magnetic Immunocaptured-Loop-Mediated Isothermal Amplification Assay |
title_fullStr |
Detection of Shigella in Milk and Clinical Samples by Magnetic Immunocaptured-Loop-Mediated Isothermal Amplification Assay |
title_full_unstemmed |
Detection of Shigella in Milk and Clinical Samples by Magnetic Immunocaptured-Loop-Mediated Isothermal Amplification Assay |
title_sort |
detection of shigella in milk and clinical samples by magnetic immunocaptured-loop-mediated isothermal amplification assay |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Microbiology |
issn |
1664-302X |
publishDate |
2018-02-01 |
description |
Shigella is an important human food-borne zoonosis bacterial pathogen, and can cause clinically severe diarrhea. There is an urgent need to develop a specific, sensitive, and rapid methodology for detection of this pathogen. In this study, loop-mediated isothermal amplification (LAMP) combined with magnetic immunocapture assay (IC-LAMP) was first developed for the detection of Shigella in pure culture, artificial milk, and clinical stool samples. This method exhibited a detection limit of 8.7 CFU/mL. Compared with polymerase chain reaction, IC-LAMP is sensitive, specific, and reliable for monitoring Shigella. Additionally, IC-LAMP is more convenient, efficient, and rapid than ordinary LAMP, as it is more efficiently enriches pathogen cells without extraction of genomic DNA. Under isothermal conditions, the amplification curves and the green fluorescence were detected within 30 min in the presence of genomic DNA template. The overall analysis time was approximately 1 h, including the enrichment and lysis of the bacterial cells, a significantly short detection time. Therefore, the IC-LAMP methodology described here is potentially useful for the efficient detection of Shigella in various samples. |
topic |
Shigella IC-LAMP immunocapture detection specificity |
url |
http://journal.frontiersin.org/article/10.3389/fmicb.2018.00094/full |
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