Pérez-Méndez A, Chandler J C, Bisha B, Goodridge L D
Department of Animal Sciences, Colorado State University, Fort Collins, CO, USA.
Department of Animal Science, University of Wyoming, Laramie, WY, USA.
J Virol Methods. 2014 Sep;206:95-8. doi: 10.1016/j.jviromet.2014.05.025. Epub 2014 Jun 6.
Detecting low concentrations of enteric viruses in water is needed for public health-related monitoring and control purposes. Thus, there is a need for sensitive, rapid and cost effective enteric viral concentration methods compatible with downstream molecular detection. Here, a virus concentration method based on adsorption of the virus to an anion exchange resin and direct isolation of nucleic acids is presented. Ten liter samples of tap water spiked with different concentrations (10-10,000 TCID50/10 L) of human adenovirus 40 (HAdV-40), hepatitis A virus (HAV) or rotavirus (RV) were concentrated and detected by real time PCR or real time RT-PCR. This method improved viral detection compared to direct testing of spiked water samples where the ΔCt was 12.1 for AdV-40 and 4.3 for HAV. Direct detection of RV in water was only possible for one of the three replicates tested (Ct of 37), but RV detection was improved using the resin method (all replicates tested positive with an average Ct of 30, n=3). The limit of detection of the method was 10 TCID50/10 L for HAdV-40 and HAV, and 100 TCID50/10 L of water for RV. These results compare favorably with detection limits reported for more expensive and laborious methods.
为了进行与公共卫生相关的监测和控制，需要检测水中低浓度的肠道病毒。因此，需要一种灵敏、快速且经济高效的肠道病毒浓缩方法，该方法要与下游分子检测兼容。在此，我们提出一种基于病毒吸附到阴离子交换树脂并直接分离核酸的病毒浓缩方法。采集了10升添加了不同浓度（10 - 10,000 TCID50/10 L）人腺病毒40（HAdV - 40）、甲型肝炎病毒（HAV）或轮状病毒（RV）的自来水样本，通过实时PCR或实时RT - PCR进行浓缩和检测。与直接检测添加病毒的水样相比，该方法提高了病毒检测能力，其中AdV - 40的ΔCt为12.1，HAV的ΔCt为4.3。对于所测试的三个重复样本中的一个，水中RV的直接检测仅在Ct值为37时才有可能，但使用树脂方法可提高RV检测能力（所有重复样本检测均为阳性，平均Ct值为30，n = 3）。该方法对HAdV - 40和HAV的检测限为10 TCID50/10 L，对RV的检测限为100 TCID50/10 L水。这些结果与报道的更昂贵且费力的方法的检测限相比具有优势。
