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RNase H2 triggered visual loop-mediated isothermal amplification combining smartphone assisted all-in-one aptamer magnetic enrichment device for ultrasensitive culture-independent detection of Salmonella Typhimurium in chicken meat

Pre-enrichment cultural continues to remain time-consuming and labor-intensive in the detection of low-concentration bacteria in meat samples. Culture-independent detection method is urgently needed in POCT technology. In order to be able to proceed with culture-independent pre-enrichment for low-co...

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Published in:Sensors and actuators. B, Chemical Chemical, 2023-04, Vol.380, p.133399, Article 133399
Main Authors: Jia, Kaiyuan, Xiao, Renhang, Lin, Qijie, Gou, Hongchao, Peng, Junhao, Liang, Yucen, Shen, Haiyan, Cai, Mingrui, Liao, Ming, Zhang, Jianmin, Han, Yuxing
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Language:English
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Summary:Pre-enrichment cultural continues to remain time-consuming and labor-intensive in the detection of low-concentration bacteria in meat samples. Culture-independent detection method is urgently needed in POCT technology. In order to be able to proceed with culture-independent pre-enrichment for low-concentration pathogen detection, a RNase H2 enzyme triggered all-in-one device-based visual Loop-mediated Isothermal Amplification detection method was developed, combining with Salmonella Typhimurium specific Aptamer Magnetic Enrichment steps, termed as the AME-LAMP method. In order to better enable the AME-LAMP method to perform on-site culture-independent detection, a palm-held all-in-one device (PAD) was designed and manufactured. Further, a self-developed smart phone fluorescence analysis app could complete end-point detection, thus eliminating miscalculation. RNase H2 Enzyme triggered AME-LAMP method could detect Salmonella as low as 5.5 CFU/mL while conventional LAMP only detected 550 CFU/mL. By verifying the detection performance of AME-LAMP methods in chicken meat samples, AME-LAMP shows the potential to achieve ultrasensitive Salmonella detection by comparing with ISO isolation method and normal LAMP. With the assistance of smart phone-based PAD, it can be reliably applied for the on-site culture-independent detection of Salmonella Typhimurium. [Display omitted] •RNase H2 triggered LAMP combining with smartphone-assisted device was developed.•The limit of Aptamer Magnetic Enrichment LAMP detection is as low as 5.5 CFU/ mL.•Palm-held all-in-one device was verified and appropriate for on-site detection.•Smart phone app was self-developed for end-point visual fluorescence detection.
ISSN:0925-4005
1873-3077
DOI:10.1016/j.snb.2023.133399