Journal
DIAGNOSTICS
Volume 11, Issue 4, Pages -Publisher
MDPI
DOI: 10.3390/diagnostics11040700
Keywords
multiplex PCR-lateral flow biosensor; point-of-care testing; Salmonella Typhi; Salmonella Paratyphi A; food handlers; typhoid carriers
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Funding
- Universiti Sains Malaysia [304/CIPPM/61310046, 1001/PSKBP/8630016, 1001/CIPPM/AUPI00256]
- Higher Institution Centre of Excellence (HICoE) Grant from Ministry of Higher Education, Malaysia [311/CIPPM/440 1005]
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The developed mPCR-LFB system allows for rapid detection of Salmonella Typhi and Salmonella Paratyphi A with high sensitivity and specificity, outperforming conventional agarose gel electrophoresis in terms of speed. Through a prospective cohort study on stool samples of 1176 food handlers, the system showed potential for rapid detection of typhoid carriers in surveillance programs.
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 10(2) 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.
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