Name: cost-effective method for microbial source tracking using specific human and animal viruses
Uploaded: 2011-12-03T14:00:00
Description: Watch this Scientific Journal Video about Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses at JoVE.com

部分支持这项工作是由西班牙政府“部教育Ÿ Ciencia”（项目AGL2008-05275-C01/ALI），由欧盟研究框架7 VIROBATHE资助项目（合同号513648），VIROCLIME（合同号243923 ）和水的加泰罗尼亚机构，通讯社Catalana艾瓜（ACA），Departament去控制我Millora dels Ecosistemes游泳。在研究开发，玛塔Rusiñol是加泰罗尼亚政府“AGAUR”（FI - DGR）的研究员。

1。病毒颗粒的浓度水样中<ol><li>水样的采集和空调<ol><li>收集最低为2 10％的样本大​​号平底塑料容器和过程控制作为一个额外的样品复制。这最后的样本将会被添加与已知的病毒颗粒量，并作为控制使用。 注意：建议有特殊独立的材料加标样品（瓶，管件，等）。 </li><li>检查校准conductimeter，如果有必要重新调整。准备一个额外的塑料10升容器中使用以前调整到足够的导电性（见下文1.6?...

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<html>描述的步骤将履行为日常的环境和公众健康实验室的拟合方法的条件是：重现性好，质量可靠，简单和成本效益。该协议是简单的，但它必须严格遵守。低电导率样品中没有添加人工海水盐的要求的浓度将大大降低病毒的复苏将的情况下，如果搅拌时间对絮凝显着降低（例如少于5小时）。 目前应用的MST工具一般都是基于分子生物学技术。由不同的群体开发的研究显示出?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>高速离心机（8000 XG） </td><td> Berckman库尔特</td><td>阿凡提J - 20XP </td><td></td></tr><tr><td> pH计，温度计和conductimeter </td><td> Afora </td><td> LPPC3003 </td><td></td></tr><tr><td>塑料管长100-200厘米</td><td> Deltalab </td><td> 350059 </td><td></td></tr><tr><td>无菌毕业于一次性移液器</td><td> Labclinics </td><td> PN10E1 </td><td></td></tr><tr><td> 1.5和10-15毫升（Eppendorf公司，猎鹰等）的无菌塑料管</td><td> Afora </td><td> KA298/00 </td><td></td></tr><tr><td>离心机壶（500毫升） </td><td> Fisher Scientific则</td><td> SE5753512 </td><td></td></tr><tr><td>磁力搅拌器和磁铁（每样本） </td><td> Fisher Scientific则</td><td> 10510 </td><td></td></tr><tr><td>平底的玻璃或塑料容器，允许使用磁力搅拌器</td><td> Deltalab </td><td> 191642 </td><td></td></tr><tr><td>一个蠕动泵取出上清液（或水喷射真空泵） </td><td>沃森的马洛</td><td> 323E / D转换</td><td></td></tr><tr><td>定时关机搅拌后8-10小时</td><td> Deltalab </td><td> 900400 </td><td></td></tr><tr><td>盐酸（1N和0.1N） </td><td> Panreac </td><td> 141020.1611 </td><td></td></tr><tr><td>氢氧化钠（1N） </td><td> Panreac </td><td> 131687.1211 </td><td></td></tr><tr><td>人工海水的海盐</td><td>西格玛</td><td> S9883 </td><td></td></tr><tr><td>脱脂牛奶（SM） </td><td> DIFCO </td><td> 232100 </td><td></td></tr><tr><td>磷酸盐缓冲液pH值75 </td><td></td><td></td><td> 1:2 V / V无菌娜2 HPO 4 0,2中号的NaH 2 PO 4 0.2，在pH 7.5中号</td></tr><tr><td>硫代</td><td> Panreac </td><td> 121879.1209 </td><td>请在水中10％的解决方案</td></tr><tr><td> QIAamp病毒RNA试剂盒</td><td> Qiagen公司</td><td> 52904 </td><td></td></tr><tr><td> 96孔光学反应板（500个单位） </td><td>应用生物系统公司</td><td> 43426659 </td><td></td></tr><tr><td>光学粘合剂覆盖（100个单位） </td><td>应用生物系统公司</td><td> 4311971 </td><td></td></tr><tr><td> TaqMan探针环境PCR扩增预混试剂2X </td><td>应用生物系统公司</td><td> 4396838 </td><td></td></tr></tbody></table>

cost-effective method for microbial source tracking using specific human and animal viruses

微生物污染的环境，代表着一个重大的健康风险。古典细菌的粪便指标显示有显著的局限性，病毒更耐许多失活过程和标准的粪便指标不告知的污染源。成本效益的方法为水和分子检测病毒浓度的发展，有利于病毒的适用性作为粪便污染的指标，并作为微生物源追踪（MST）工具。腺病毒，多瘤病毒是DNA病毒感染具体包括人类在内的脊椎动物物种，并坚持在粪便和/或研究所有地区的尿液排出体外。在以往的研究中，我们建议人类腺病毒（HAdV）和JC多瘤病毒定量PCR（qPCR）（JCPyV）作为人类粪便污染指数的量化。最近，我们已经制定了具体量化的POR qPCR实验电影的腺病毒（反对家庭暴力伙伴关系）和牛多瘤病毒（BPyV）每1-10试管基因组副本的敏感动物粪便污染的标记。在这项研究中，我们目前所应遵循的程序，以确定在使用这些工具的水样本的污染来源。作为代表性的结果为例，提出高水平的硝酸盐在地下水的病毒分析显示。 低或中度污染水域的病毒检测需要从几升的水至少到一个更小的体积，一个过程，通常包括两个系列的浓缩步骤的病毒浓度。这有些繁琐的程序，观察病毒回收率的变异显著妨碍同时处理大量的水样。 为了消除由两个步骤的程序所造成的瓶颈，我们已经申请了一个一步到位的协议，在以前的studieS和适用于水基质的多样性。该程序包括：10升的水样酸化，絮凝脱脂奶，重力沉淀絮凝材料，沉淀，离心收集，在磷酸盐缓冲液10毫升沉淀再悬浮。病毒集中用于提取病毒核酸和具体利益的腺病毒和多瘤病毒是由qPCR的量化。使用这种低成本的浓度的方法，可同时分析大量样品。 该程序已应用于洗浴水，海水和河水的分析，在这项研究中，我们目前的地下水样品分析的结果。这种高通量的定量方法是可靠的，简单，成本效益。

Watch this Scientific Journal Video about Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses at JoVE.com

Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses

这项研究描述为粪/尿污染或使用人/猪/牛的DNA病毒，腺病毒和提出的MST工具多瘤病毒，具体量化的qPCR在水中的硝酸盐污染的来源识别成本效益的方法。

微生物源使用特定的人类和动物病毒的追踪成本效益的方法

Microbial contamination of the environment represents a significant health risk. Classical bacterial fecal indicators have shown to have significant ...

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