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Proceedings Paper

Point of care nucleic acid detection of viable pathogenic bacteria with isothermal RNA amplification based paper biosensor
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Paper Abstract

Food-borne pathogens such as Listeria monocytogenes have been recognized as a major cause of human infections worldwide, leading to substantial health problems. Food-borne pathogen identification needs to be simpler, cheaper and more reliable than the current traditional methods. Here, we have constructed a low-cost paper biosensor for the detection of viable pathogenic bacteria with the naked eye. In this study, an effective isothermal amplification method was used to amplify the hlyA mRNA gene, a specific RNA marker in Listeria monocytogenes. The amplification products were applied to the paper biosensor to perform a visual test, in which endpoint detection was performed using sandwich hybridization assays. When the RNA products migrated along the paper biosensor by capillary action, the gold nanoparticles accumulated at the designated Test line and Control line. Under optimized experimental conditions, as little as 0.5 pg/μL genomic RNA from Listeria monocytogenes could be detected. The whole assay process, including RNA extraction, amplification, and visualization, can be completed within several hours. The developed method is suitable for point-of-care applications to detect food-borne pathogens, as it can effectively overcome the false-positive results caused by amplifying nonviable Listeria monocytogenes.

Paper Details

Date Published: 17 September 2014
PDF: 9 pages
Proc. SPIE 9230, Twelfth International Conference on Photonics and Imaging in Biology and Medicine (PIBM 2014), 923007 (17 September 2014); doi: 10.1117/12.2068649
Show Author Affiliations
Hongxing Liu, South China Normal Univ. (China)
Da Xing, South China Normal Univ. (China)
Xiaoming Zhou, South China Normal Univ. (China)

Published in SPIE Proceedings Vol. 9230:
Twelfth International Conference on Photonics and Imaging in Biology and Medicine (PIBM 2014)
Qingming Luo; Lihong V. Wang; Valery V. Tuchin, Editor(s)

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