Nucleic Acid-Based Lateral Flow Biosensor for <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi: A Detection in Stool Samples of Suspected Carriers
A multiplex rapid detection system, based on a PCR-lateral flow biosensor (mPCR-LFB) was developed to identify <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi A from suspected carriers. The lower detection limit for <i>S</i>. Typhi and <i>S</i>...
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doaj-2ffe0a32fac44ad8b214fabfffa91a882021-04-14T23:02:46ZengMDPI AGDiagnostics2075-44182021-04-011170070010.3390/diagnostics11040700Nucleic Acid-Based Lateral Flow Biosensor for <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi: A Detection in Stool Samples of Suspected CarriersZulkiply Nor Amalina0Muhammad Fazli Khalid1Sjafri Faizul Rahman2Muhamad Nuramin Ahmad3Mohamad Ahmad Najib4Asma Ismail5Ismail Aziah6Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, MalaysiaA multiplex rapid detection system, based on a PCR-lateral flow biosensor (mPCR-LFB) was developed to identify <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi A from suspected carriers. The lower detection limit for <i>S</i>. Typhi and <i>S</i>. Paratyphi A was 0.16 and 0.08 ng DNA equivalent to 10 and 10<sup>2</sup> CFU/mL, respectively. Lateral flow biosensor was used for visual detection of mPCR amplicons (stgA, SPAint, ompC, internal amplification control) by labeling forward primers with fluorescein-isothiocyanate (FITC), Texas Red, dinitrophenol (DNP) and digoxigenin (DIG) and reverse primers with biotin. Binding of streptavidin-colloidal gold conjugate with the amplicons resulted in formation of a red color dots on the strip after 15–20 min of sample exposure. The nucleic acid lateral flow analysis of the mPCR-LFB was better in sensitivity and more rapid than the conventional agarose gel electrophoresis. Moreover, the mPCR-LFB showed 100% sensitivity and specificity when evaluated with stools spiked with 100 isolates of <i>Salmonella</i> genus and other bacteria. A prospective cohort study on stool samples of 1176 food handlers in outbreak areas (suspected carriers) resulted in 23 (2%) positive for <i>S</i>. Typhi. The developed assay has potential to be used for rapid detection of typhoid carriers in surveillance program.https://www.mdpi.com/2075-4418/11/4/700multiplex PCR-lateral flow biosensorpoint-of-care testing<i>Salmonella</i> Typhi<i>Salmonella</i> Paratyphi Afood handlerstyphoid carriers |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Zulkiply Nor Amalina Muhammad Fazli Khalid Sjafri Faizul Rahman Muhamad Nuramin Ahmad Mohamad Ahmad Najib Asma Ismail Ismail Aziah |
spellingShingle |
Zulkiply Nor Amalina Muhammad Fazli Khalid Sjafri Faizul Rahman Muhamad Nuramin Ahmad Mohamad Ahmad Najib Asma Ismail Ismail Aziah Nucleic Acid-Based Lateral Flow Biosensor for <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi: A Detection in Stool Samples of Suspected Carriers Diagnostics multiplex PCR-lateral flow biosensor point-of-care testing <i>Salmonella</i> Typhi <i>Salmonella</i> Paratyphi A food handlers typhoid carriers |
author_facet |
Zulkiply Nor Amalina Muhammad Fazli Khalid Sjafri Faizul Rahman Muhamad Nuramin Ahmad Mohamad Ahmad Najib Asma Ismail Ismail Aziah |
author_sort |
Zulkiply Nor Amalina |
title |
Nucleic Acid-Based Lateral Flow Biosensor for <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi: A Detection in Stool Samples of Suspected Carriers |
title_short |
Nucleic Acid-Based Lateral Flow Biosensor for <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi: A Detection in Stool Samples of Suspected Carriers |
title_full |
Nucleic Acid-Based Lateral Flow Biosensor for <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi: A Detection in Stool Samples of Suspected Carriers |
title_fullStr |
Nucleic Acid-Based Lateral Flow Biosensor for <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi: A Detection in Stool Samples of Suspected Carriers |
title_full_unstemmed |
Nucleic Acid-Based Lateral Flow Biosensor for <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi: A Detection in Stool Samples of Suspected Carriers |
title_sort |
nucleic acid-based lateral flow biosensor for <i>salmonella</i> typhi and <i>salmonella</i> paratyphi: a detection in stool samples of suspected carriers |
publisher |
MDPI AG |
series |
Diagnostics |
issn |
2075-4418 |
publishDate |
2021-04-01 |
description |
A multiplex rapid detection system, based on a PCR-lateral flow biosensor (mPCR-LFB) was developed to identify <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi A from suspected carriers. The lower detection limit for <i>S</i>. Typhi and <i>S</i>. Paratyphi A was 0.16 and 0.08 ng DNA equivalent to 10 and 10<sup>2</sup> CFU/mL, respectively. Lateral flow biosensor was used for visual detection of mPCR amplicons (stgA, SPAint, ompC, internal amplification control) by labeling forward primers with fluorescein-isothiocyanate (FITC), Texas Red, dinitrophenol (DNP) and digoxigenin (DIG) and reverse primers with biotin. Binding of streptavidin-colloidal gold conjugate with the amplicons resulted in formation of a red color dots on the strip after 15–20 min of sample exposure. The nucleic acid lateral flow analysis of the mPCR-LFB was better in sensitivity and more rapid than the conventional agarose gel electrophoresis. Moreover, the mPCR-LFB showed 100% sensitivity and specificity when evaluated with stools spiked with 100 isolates of <i>Salmonella</i> genus and other bacteria. A prospective cohort study on stool samples of 1176 food handlers in outbreak areas (suspected carriers) resulted in 23 (2%) positive for <i>S</i>. Typhi. The developed assay has potential to be used for rapid detection of typhoid carriers in surveillance program. |
topic |
multiplex PCR-lateral flow biosensor point-of-care testing <i>Salmonella</i> Typhi <i>Salmonella</i> Paratyphi A food handlers typhoid carriers |
url |
https://www.mdpi.com/2075-4418/11/4/700 |
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