Simultaneous detection of multiple bacterial and viral aquatic pathogens using a fluorogenic loop‐mediated isothermal amplification‐based dual‐sample microfluidic chip

2020 ◽  
Author(s):  
Qian‐Jin Zhou ◽  
Jian‐Fei Lu ◽  
Xiu‐Rong Su ◽  
Jing‐Lei Jin ◽  
Shang‐Yang Li ◽  
...  
Keyword(s):  
Microfluidic Chip ◽  
Simultaneous Detection ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Aquatic Pathogens

scholarly journals Fast, Precise, and Reliable Multiplex Detection of Potato Viruses by Loop-Mediated Isothermal Amplification

2020 ◽  
Vol 21 (22) ◽  
pp. 8741
Author(s):  
Güven Edgü ◽  
Lena Julie Freund ◽  
Stefanie Hartje ◽  
Eckhard Tacke ◽  
Hans-Reinhard Hofferbert ◽  
...  
Keyword(s):  
Disease Management ◽  
Management Practices ◽  
Tobacco Rattle Virus ◽  
Simultaneous Detection ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Tuber Quality ◽  
Loop Mediated Isothermal Amplification ◽  
Tuber Necrosis ◽  
Corky Ringspot

Potato is an important staple food crop in both developed and developing countries. However, potato plants are susceptible to several economically important viruses that reduce yields by up to 50% and affect tuber quality. One of the major threats is corky ringspot, which is a tuber necrosis caused by tobacco rattle virus (TRV). The appearance of corky ringspot symptoms on tubers prior to commercialization results in ≈ 45% of the tubers being downgraded in quality and value, while ≈ 55% are declared unsaleable. To improve current disease management practices, we have developed simple diagnostic methods for the reliable detection of TRV without RNA purification, involving minimalized sample handling (mini), subsequent improved colorimetric loop-mediated isothermal amplification (LAMP), and final verification by lateral-flow dipstick (LFD) analysis. Having optimized the mini-LAMP-LFD approach for the sensitive and specific detection of TRV, we confirmed the reliability and robustness of this approach by the simultaneous detection of TRV and other harmful viruses in duplex LAMP reactions. Therefore, our new approach offers breeders, producers, and farmers an inexpensive and efficient new platform for disease management in potato breeding and cultivation.

scholarly journals Rapid and simultaneous detection of Campylobacter spp. and Salmonella spp. in chicken samples by duplex loop-mediated isothermal amplification coupled with a lateral flow biosensor assay

PLoS ONE ◽  
2021 ◽  
Vol 16 (7) ◽  
pp. e0254029
Author(s):  
Thanawat Sridapan ◽  
Wanida Tangkawsakul ◽  
Tavan Janvilisri ◽  
Taradon Luangtongkum ◽  
Wansika Kiatpathomchai ◽  
...  
Keyword(s):  
Simultaneous Detection ◽  
Chicken Meat ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Salmonella Spp ◽  
Promising Alternative ◽  
Loop Mediated Isothermal Amplification ◽  
Alternative Approach ◽  
Meat Samples ◽  
Sensitivity Specificity

Development of a simple, rapid and specific assay for the simultaneous detection of Campylobacter spp. and Salmonella spp. based on duplex loop-mediated isothermal amplification (d-LAMP), combined with lateral-flow biosensor (LFB) is reported herein. LAMP amplicons of both pathogens were simultaneously amplified and specifically differentiated by LFB. The specificity of the d-LAMP-LFB was evaluated using a set of 68 target and 12 non-target strains, showing 100% inclusivity and exclusivity. The assay can simultaneously detect Campylobacter and Salmonella strains as low as 1 ng and 100 pg genomic DNA per reaction, respectively. The lowest inoculated detection limits for Campylobacter and Salmonella species in artificially contaminated chicken meat samples were 103 CFU and 1 CFU per 25 grams, respectively, after enrichment for 24 h. Furthermore, compared to culture-based methods using field chicken meat samples, the sensitivity, specificity and accuracy of d-LAMP- LFB were 95.6% (95% CI, 78.0%-99.8%), 71.4% (95% CI, 29.0%-96.3%) and 90.0% (95% CI, 73.4%-97.8%), respectively. The developed d-LAMP-LFB assay herein shows great potentials for the simultaneous detection of the Campylobacter and Salmonella spp. and poses a promising alternative approach for detection of both pathogens with applications in food products.

Loop-mediated isothermal amplification-based microfluidic chip for pathogen detection

2018 ◽  
Vol 60 (2) ◽  
pp. 201-224 ◽  
Author(s):  
Yuting Shang ◽  
Jiadi Sun ◽  
Yongli Ye ◽  
Jumei Zhang ◽  
Yinzhi Zhang ◽  
...  
Keyword(s):  
Microfluidic Chip ◽  
Pathogen Detection ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification

scholarly journals Fast and Parallel Detection of Four Ebola Virus Species on a Microfluidic-Chip-Based Portable Reverse Transcription Loop-Mediated Isothermal Amplification System

Micromachines ◽  
2019 ◽  
Vol 10 (11) ◽  
pp. 777 ◽  
Author(s):  
Xue Lin ◽  
Xiangyu Jin ◽  
Bin Xu ◽  
Ruliang Wang ◽  
Rongxin Fu ◽  
...  
Keyword(s):  
Microfluidic Chip ◽  
Reverse Transcription ◽  
Ebola Virus ◽  
Limit Of Detection ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Virus Species ◽  
Fluorescence Detector ◽  
Parallel Detection ◽  
Loop Mediated Isothermal Amplification

Considering the lack of official vaccines and medicines for Ebola virus infection, reliable diagnostic methods are necessary for the control of the outbreak and the spread of the disease. We developed a microfluidic-chip-based portable system for fast and parallel detection of four Ebola virus species. The system is based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and consists of four specific LAMP primers, a disc microfluidic chip, and a portable real-time fluorescence detector. It could specifically and parallelly distinguish four species of the Ebola virus after only one sampling, including the Zaire Ebola virus, the Sudan Ebola virus, the Bundibugyo Ebola virus, and the Tai Forest Ebola virus, without cross-contamination. The limit of detection was as small as 10 copies per reaction, while the total consumption of sample and reagent was 0.94 μL per reaction. The final results could be obtained in 50 min after one addition of sample and reagent mixture. This approach provides simplicity, high sensitivity, and multi-target parallel detection at a low cost, which could enable convenient and effective on-site detections of the Ebola virus in the outdoors, remote areas, and modern hospitals.

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