核酸标准物质在金针菇中坂崎克罗诺杆菌（Cronobacter spp.）快速检测中的应用,Journal of Food Quality

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核酸标准物质在金针菇中坂崎克罗诺杆菌（Cronobacter spp.）快速检测中的应用
Journal of Food Quality ( IF 3.3 ) Pub Date : 2020-11-17 , DOI: 10.1155/2020/8821330
Huandie Yang _{1,

2} , Danyang Guo ₁ , Leilei Chen ₁ , Alexander Suvorov ₁ , Qingxin Zhou ₁ , Junhua Wang ₁ , Xiaofeng Gao ₃ , Jinyu Yang ₁ , Xiangyan Chen ₁

Affiliation

坂崎克罗诺杆菌（C. sakazakii）广泛存在于环境中，是一种常见的食源性病原微生物。对免疫力低下的婴幼儿和老年人可引起脑膜炎、菌血症和坏死性小肠结肠炎。因此，开发食源性阪崎肠杆菌快速检测方法对食品安全控制具有重要意义。本研究开发了一种基因组核酸参考物质，作为快速检测食用菌金针菇中阪崎肠杆菌的阳性参考物质。首先，从阪崎肠杆菌标准菌株中提取基因组核酸，并将核酸冻干。冻干粉的均匀度测试结果表明，样品之间没有显着差异。在4℃和37℃14天的短期稳定性试验中，核酸对照品性能良好，在4℃和-20℃8个月的长期稳定性试验中，核酸对照品的性质良好。样品还是很稳定的。为验证该核酸对照品的应用，将其作为PCR检测的阳性对照模板，应用于F. velutipes中阪崎肠杆菌的快速检测。发现 15 个 F. velutipes 样本中有 13 个对 C. sakazakii 呈阳性。据我们所知，这是首次尝试以核酸参照物为阳性参照物，采用PCR方法检测F. velutipes中的C. sakazakii。本研究有助于推动核酸参比物质在食源性病原体检测中的广泛深入应用。核酸对照品性质良好，在4℃和-20℃8个月的长期稳定性试验中，样品仍然稳定。为验证该核酸对照品的应用，将其作为PCR检测的阳性对照模板，应用于F. velutipes中阪崎肠杆菌的快速检测。发现 15 个 F. velutipes 样本中有 13 个对 C. sakazakii 呈阳性。据我们所知，这是首次尝试以核酸参照物为阳性参照物，采用PCR方法检测F. velutipes中的C. sakazakii。本研究有助于推动核酸参比物质在食源性病原体检测中的广泛深入应用。核酸对照品性质良好，在4℃和-20℃8个月的长期稳定性试验中，样品仍然稳定。为验证该核酸对照品的应用，将其作为PCR检测的阳性对照模板，应用于F. velutipes中阪崎肠杆菌的快速检测。发现 15 个 F. velutipes 样本中有 13 个对 C. sakazakii 呈阳性。据我们所知，这是首次尝试以核酸参照物为阳性参照物，采用PCR方法检测F. velutipes中的C. sakazakii。本研究有助于推动核酸参比物质在食源性病原体检测中的广泛深入应用。样品还是很稳定的。为验证该核酸对照品的应用，将其作为PCR检测的阳性对照模板，应用于F. velutipes中阪崎肠杆菌的快速检测。发现 15 个 F. velutipes 样本中有 13 个对 C. sakazakii 呈阳性。据我们所知，这是首次尝试以核酸参照物为阳性参照物，采用PCR方法检测F. velutipes中的C. sakazakii。本研究有助于推动核酸参比物质在食源性病原体检测中的广泛深入应用。样品还是很稳定的。为验证该核酸对照品的应用，将其作为PCR检测的阳性对照模板，应用于F. velutipes中阪崎肠杆菌的快速检测。发现 15 个 F. velutipes 样本中有 13 个对 C. sakazakii 呈阳性。据我们所知，这是首次尝试以核酸参照物为阳性参照物，采用PCR方法检测F. velutipes中的C. sakazakii。本研究有助于推动核酸参比物质在食源性病原体检测中的广泛深入应用。velutipes 样品对 C. sakazakii 呈阳性。据我们所知，这是首次尝试以核酸参照物为阳性参照物，采用PCR方法检测F. velutipes中的C. sakazakii。本研究有助于推动核酸参比物质在食源性病原体检测中的广泛深入应用。velutipes 样品对 C. sakazakii 呈阳性。据我们所知，这是首次尝试以核酸参照物为阳性参照物，采用PCR方法检测F. velutipes中的C. sakazakii。本研究有助于推动核酸参比物质在食源性病原体检测中的广泛深入应用。

"点击查看英文标题和摘要"

Application of Nucleic Acid Reference Material for Rapid Detection of Cronobacter sakazakii (Cronobacter spp.) in Flammulina velutipes

Cronobacter sakazakii (C. sakazakii) widely exists in the environment and is a common foodborne pathogenic microorganism. It can cause meningitis, bacteremia, and necrotizing enterocolitis in infants and elderly people with low immunity. Therefore, the development of the rapid detection method for foodborne pathogen C. sakazakii is very important for food safety control. This study has developed a kind of genomic nucleic acid reference material as a positive reference for the rapid detection of C. sakazakii in the edible fungus Flammulina velutipes. First, the genomic nucleic acids were extracted from the standard strain of C. sakazakii, and the nucleic acids were lyophilized. The uniformity test result of the lyophilized powder showed that there were no significant differences between samples. In short-term stability tests at 4°C and 37°C for 14 days, the properties of the nucleic acid reference material were good, and in long-term stability tests at 4°C and −20°C for 8 months, the samples are still stable. To verify the application of this nucleic acid reference material, it was used as a positive control template for PCR detection and applied to the rapid detection of C. sakazakii in F. velutipes. It was found that 13 of the 15 F. velutipes samples were positive for C. sakazakii. As far as we know, this is the first attempt to detect C. sakazakii in F. velutipes using the PCR method with nucleic acid reference material as the positive reference. This study is helpful in promoting the extensive and in-depth application of nucleic acid reference materials in foodborne pathogens detection.

更新日期：2020-11-17

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