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

Hongxing Liu; Da Xing; Xiaoming Zhou

doi:10.1117/12.2068649

17 September 2014 Point of care nucleic acid detection of viable pathogenic bacteria with isothermal RNA amplification based paper biosensor

Hongxing Liu, Da Xing, Xiaoming Zhou

Author Affiliations +

Proceedings Volume 9230, Twelfth International Conference on Photonics and Imaging in Biology and Medicine (PIBM 2014); 923007 (2014) https://doi.org/10.1117/12.2068649
Event: Twelfth International Conference on Photonics and Imaging in Biology and Medicine (PIBM 2014), 2014, Wuhan, China

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.

Citation Download Citation

Hongxing Liu, Da Xing, and Xiaoming Zhou "Point of care nucleic acid detection of viable pathogenic bacteria with isothermal RNA amplification based paper biosensor", Proc. SPIE 9230, Twelfth International Conference on Photonics and Imaging in Biology and Medicine (PIBM 2014), 923007 (17 September 2014); https://doi.org/10.1117/12.2068649

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