期刊
ACS SENSORS
卷 3, 期 2, 页码 403-409出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.7b00825
关键词
nucleic-acid based tests; paper; microfluidics; infectious disease; diagnosis
资金
- University of Glasgow Fellowship
- BBSRC [Bb/L00464x/1]
- DBT [BT/IN/Indo-UK/FADH/43/PC/2013]
- ERC [340117]
- EPSRC [EP/K027611/1]
- College of Science and Engineering Scholarship from the University of Glasgow
- BBSRC [BB/L00464X/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/L00464X/1] Funding Source: researchfish
- Engineering and Physical Sciences Research Council [EP/K027611/1] Funding Source: researchfish
The health and well-being of cattle is an important issue in maintaining and increasing global agricultural output. In dairy production within low and middle income countries (LMICs), there is a significant biosensing challenge in detecting sexually transmitted infection (STI) pathogens during animal husbandry, due in part to difficulties associated with the limited infrastructure for veterinary medicine. Here we demonstrate low-cost, multiplexed, and sample-to-answer paper-origami tests for the detection of three bovine infectious reproductive diseases in semen samples, collected at a test site in rural India. Pathogen DNA from one viral pathogen, bovine herpes virus-1 (BoHV-1), and two bacteria (Brucella and Leptospira) was extracted, amplified (using loop-mediated isothermal amplification, LAMP), and detected fluorescently, enabling <1 pg (similar to from 115 to 274 copies per reaction) of target genomic DNA to be measured. Data was collected as a fluorescence signal either visually, using a low-cost hand-held torch, or digitally with a mobile-phone camera. Limits of detection and sensitivities of the paper-origami device for the three pathogens were also evaluated using pathogen-inoculated semen samples and were as few as 50 Leptospira organisms, 50 CFU Brucella, and I TCID50 BoHV(-1). Semen samples from elite bulls at a germplasm center were also tested in double-blind tests, as a demonstrator for a low-cost, user-friendly point-of-care sensing platform, for in-the-field resource-limited regions. The sensors showed excellent levels of sensitivity and specificity, and for the first time a demonstrated ability of the application of paper microfluidics devices for the diagnosis multiple infectious diseases from semen samples.
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