一种用于检测灌溉水中烟草花叶病毒的浓缩方法的开发。
Development of a concentration method for detection of tobacco mosaic virus in irrigation water.
作者信息
Chen Wei, Liu Wenting, Jiao Honghong, Zhang Huawei, Cheng Julong, Wu Yunfeng
机构信息
State Key Laboratory of Crop Stress Biology for Arid Areas and Key Laboratory of Crop Pest Integrated Pest Management on the Loess Plateau of Ministry of Agriculture, College of Plant Protection, Northwest A&F University, Yangling, 712100, China.
出版信息
Virol Sin. 2014 Jun;29(3):155-61. doi: 10.1007/s12250-014-3461-7. Epub 2014 Jun 18.
Tobacco mosaic virus (TMV) causes significant yield loss in susceptible crops irrigated with contaminated water. However, detection of TMV in water is difficult owing to extremely low concentrations of the virus. Here, we developed a simple method for the detection and quantification of TMV in irrigation water. TMV was reliably detected at concentrations as low as 10 viral copies/μL with real-time PCR. The sensitivity of detection was further improved using polyethylene glycol 6000 (PEG6000, MW 6000) to concentrate TMV from water samples. Among the 28 samples from Shaanxi Province examined with our method, 17 were tested positive after virus concentration. Infectivity of TMV in the original water sample as well as after concentration was confirmed using PCR. The limiting concentration of TMV in water to re-infect plants was determined as 10(2) viral copies/mL. The method developed in this study offers a novel approach to detect TMV in irrigation water, and may provide an effective tool to control crop infection.
烟草花叶病毒（TMV）会使使用受污染水灌溉的易感作物大幅减产。然而，由于水中病毒浓度极低，难以对水中的TMV进行检测。在此，我们开发了一种用于检测和定量灌溉水中TMV的简单方法。通过实时聚合酶链反应（PCR）能够可靠地检测到低至10个病毒拷贝/微升浓度的TMV。使用聚乙二醇6000（PEG6000，分子量6000）从水样中浓缩TMV，检测灵敏度进一步提高。在用我们的方法检测的来自陕西省的28个样本中，17个样本在病毒浓缩后检测呈阳性。通过PCR确认了原始水样以及浓缩后TMV的感染性。确定水中TMV再次感染植物的极限浓度为10²个病毒拷贝/毫升。本研究中开发的方法为检测灌溉水中的TMV提供了一种新途径，可能为控制作物感染提供有效工具。
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