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Nucleic Acid-Based Lateral Flow Biosensor for Salmonella Typhi and Salmonella Paratyphi: A Detection in Stool Samples of Suspected Carriers.


ABSTRACT: A multiplex rapid detection system, based on a PCR-lateral flow biosensor (mPCR-LFB) was developed to identify Salmonella Typhi and Salmonella Paratyphi A from suspected carriers. The lower detection limit for S. Typhi and S. Paratyphi A was 0.16 and 0.08 ng DNA equivalent to 10 and 102 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 Salmonella 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 S. Typhi. The developed assay has potential to be used for rapid detection of typhoid carriers in surveillance program.

SUBMITTER: Amalina ZN 

PROVIDER: S-EPMC8070779 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature

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Nucleic Acid-Based Lateral Flow Biosensor for <i>Salmonella</i> Typhi and <i>Salmonella</i> Paratyphi: A Detection in Stool Samples of Suspected Carriers.

Amalina Zulkiply Nor ZN   Khalid Muhammad Fazli MF   Rahman Sjafri Faizul SF   Ahmad Muhamad Nuramin MN   Ahmad Najib Mohamad M   Ismail Asma A   Aziah Ismail I  

Diagnostics (Basel, Switzerland) 20210414 4


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  ...[more]

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