Colorimetric Detection of Norovirus in Oyster Samples through DNAzyme as a Signaling Probe
Colorimetric Detection of Norovirus in Oyster Samples through DNAzyme as a Signaling Probe
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DOI：
10.1021/acs.jafc.7b05289
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发表时间：
2018-03-21
影响因子：
6.1
通讯作者：
Kim, Min-Gon
中科院分区：
文献类型：
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作者：
Batule, Bhagwan S.;Kim, Seong U.;Kim, Min-Gon
在世界范围内，诺如病毒是急性胃肠炎最相关的原因之一，每年导致发展中国家近 50 000 名儿童死亡。因此，迫切需要开发一种快速、低成本、准确的食源性诺如病毒感染检测方法，以降低诺如病毒造成的死亡率。考虑到诺如病毒的重要性，我们展示了一种高度灵敏和特异的比色检测方法，用于分析牡蛎样本中的人类诺如病毒基因组 I 和 II（HuNoV GI 和 GII）。这是第一份采用比色 HRPzyme 集成聚合酶链反应 (PCR) 从真实贝类样本中直接检测诺如病毒的报告。我们发现 HRPzyme 集成 PCR 方法比凝胶电泳方法更灵敏，可以检测高达 1 拷贝/mL 的 HuNoV GI 和 GII 基因组。该方法的特异性在 HuNoV GI 和 GII 中得到了成功证明。此外，我们对加标牡蛎样本中的 HuNoV 进行了检测，HRPzyme 集成 PCR 方法被证明是检测真实样本中 HuNoV 的超灵敏和选择性方法。通过将所提出的方法与便携式PCR仪相结合，通过在生产现场检测不同类型的贝类（例如牡蛎和贻贝）中的HuNoV来提高食品安全将更加可靠。
Worldwide, norovirus is one of the most associated causes of acute gastroenteritis, which leads to nearly 50 000 child deaths every year in developing countries. Therefore, there is great demand to develop a rapid, low-cost, and accurate detection assay for the foodborne norovirus infection to reduce mortality caused by norovirus. Considering the importance of norovirus, we have demonstrated a highly sensitive and specific colorimetric detection method for analysis of human norovirus genogroups I and II (HuNoV GI and GII) in oyster samples. This is the first report to employ colorimetric HRPzyme-integrated polymerase chain reaction (PCR) for direct norovirus detection from the real shellfish samples. We found that the HRPzyme-integrated PCR method is more sensitive than the gel electrophoresis approach and could detect the HuNoV GI and GII genome up to 1 copy/mL. The specificity of the proposed method was successfully demonstrated for HuNoV GI and GII. Further, we performed testing HuNoVs in the spiked oyster samples, and the HRPzyme-integrated PCR method proved to be an ultrasensitive and selective method for detecting HuNoVs in the real samples. By integration of the proposed method with the portable PCR machine, it would be more reliable to improve food safety by detecting HuNoVs in the different types of shellfish, such as oyster and mussel, at the production field.