The establishment of isothermal amplification and lateral flow device for detection of salmonella typhi and salmonella paratyphi A
Loading...
Date
2020-11
Authors
Hanaphi, Roziana Mohamed
Journal Title
Journal ISSN
Volume Title
Publisher
Pusat Pengajian Sains Perubatan, Universiti Sains Malaysia
Abstract
specificity for both targeted bacteria detection on DNA of bacteria isolates of 30 S. Typhi, 30 S. Paratyphi A, 30 Salmonella serovars and 30 non-Salmonella bacteria. The limit of detection of the CPA for S. Typhi assay was 1 fg of genomic templates and 101 CFU/ml in bacteria culture, while, the CPA for S. Paratyphi A assay was 10 fg of genomic templates and 103 CFU/ml in bacteria culture. The results obtained were consistent for both S. Typhi and S. Paratyphi A assays for the detection method with agarose gel electrophoresis (AGE) and LFA. The sensitivity and specificity of both assays were effective when tested with 54 spiked-stool samples, 56 water samples from endemic areas and three stool samples of suspected carriers. The final form of lyophilised CPA reagents was obtained provided ready-to-use reagents together with lateral flow strips for limited resource settings and transportation without a cold chain. The stability testing of both assays in different storage temperatures of 4°C, 25°C, 37°C and 45°C revealed the shelf-life of three to four months when tested in 30 days storage. As a conclusion, the CPA was effective in the detection of S. Typhi and S. Paratyphi A, which was ideal for limited-resource settings with rapid detection, simple procedure and equipment, and easy readout of the assay results using LFA.
Description
Keywords
Salmonella infections