Rapid Veterinary Diagnosis of Bovine Reproductive Infectious Diseases from Semen Using Paper-Origami DNA Microfluidics.
Rapid Veterinary Diagnosis of Bovine Reproductive Infectious Diseases from Semen Using Paper-Origami DNA Microfluidics.
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DOI：
10.1021/acssensors.7b00825
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发表时间：
2018-02
期刊：
影响因子：
8.9
通讯作者：
Zhugen Yang;Gaolian Xu;J. Reboud;S. A. Ali;G. Kaur;J. McGiven;Nongthombam Boby;P. Gupta;P. Chaudhuri;J. Cooper
中科院分区：
文献类型：
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作者：
Zhugen Yang;Gaolian Xu;J. Reboud;S. A. Ali;G. Kaur;J. McGiven;Nongthombam Boby;P. Gupta;P. Chaudhuri;J. Cooper
牛的健康和福祉是维持和增加全球农业产量的一个重要问题。在低收入和中等收入国家的乳制品生产中，在畜牧业期间检测性传播感染(STI)病原体方面存在重大的生物传感挑战，部分原因是与兽医基础设施有限有关的困难。在这里，我们展示了一种低成本、多路和样本到答案的纸质折纸测试，用于检测在印度农村一个测试地点收集的精液样本中的三种牛传染性生殖疾病。从牛疱疹病毒-1(BoHV-1)和布鲁氏菌(Brucella)和钩端螺旋体(Leptspira)两种细菌中提取病原体DNA，利用环介导的等温扩增(LAMP)进行扩增，并进行荧光检测，从而能够测量<；1pg(每次反应的∼从115到274个拷贝)。数据是以荧光信号的形式收集的，要么是用低成本的手持手电筒进行视觉收集，要么是用手机相机进行数字采集。使用接种病原体的精液样本也评估了纸折纸装置对这三种病原体的检测限和灵敏度，最低可检测到50种钩端螺旋体、50种布鲁氏菌和1种TCID50 BoHV-1。来自种质中心精英公牛的精液样本也进行了双盲测试，作为一个低成本、用户友好的护理点传感平台的演示，适用于田间资源有限的地区。传感器表现出极高的灵敏度和特异度，并首次展示了应用纸质微流控设备从精液样本中诊断多种传染病的能力。
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 (∼ 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 1 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.