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 f...

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Main Authors: Huandie Yang, Danyang Guo, Leilei Chen, Alexander Suvorov, Qingxin Zhou, Junhua Wang, Xiaofeng Gao, Jinyu Yang, Xiangyan Chen
Format: Article
Language:English
Published: Wiley 2020-01-01
Series:Journal of Food Quality
Online Access:http://dx.doi.org/10.1155/2020/8821330
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author Huandie Yang
Danyang Guo
Leilei Chen
Alexander Suvorov
Qingxin Zhou
Junhua Wang
Xiaofeng Gao
Jinyu Yang
Xiangyan Chen
author_facet Huandie Yang
Danyang Guo
Leilei Chen
Alexander Suvorov
Qingxin Zhou
Junhua Wang
Xiaofeng Gao
Jinyu Yang
Xiangyan Chen
author_sort Huandie Yang
collection DOAJ
description 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.
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spelling doaj-art-b67fa82c19b14b53acd94a20bbe452bb2025-02-03T06:46:16ZengWileyJournal of Food Quality0146-94281745-45572020-01-01202010.1155/2020/88213308821330Application of Nucleic Acid Reference Material for Rapid Detection of Cronobacter sakazakii (Cronobacter spp.) in Flammulina velutipesHuandie Yang0Danyang Guo1Leilei Chen2Alexander Suvorov3Qingxin Zhou4Junhua Wang5Xiaofeng Gao6Jinyu Yang7Xiangyan Chen8Institute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences, Key Laboratory of Agro-Products Processing Technology of Shandong Province, Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture and Rural Affairs, Jinan 250100, ChinaInstitute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences, Key Laboratory of Agro-Products Processing Technology of Shandong Province, Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture and Rural Affairs, Jinan 250100, ChinaInstitute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences, Key Laboratory of Agro-Products Processing Technology of Shandong Province, Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture and Rural Affairs, Jinan 250100, ChinaInstitute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences, Key Laboratory of Agro-Products Processing Technology of Shandong Province, Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture and Rural Affairs, Jinan 250100, ChinaInstitute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences, Key Laboratory of Agro-Products Processing Technology of Shandong Province, Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture and Rural Affairs, Jinan 250100, ChinaInstitute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences, Key Laboratory of Agro-Products Processing Technology of Shandong Province, Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture and Rural Affairs, Jinan 250100, ChinaNational R & D Center for Edible Fungus Processing Technology, Henan University, Kaifeng 475004, ChinaInstitute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences, Key Laboratory of Agro-Products Processing Technology of Shandong Province, Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture and Rural Affairs, Jinan 250100, ChinaInstitute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences, Key Laboratory of Agro-Products Processing Technology of Shandong Province, Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture and Rural Affairs, Jinan 250100, ChinaCronobacter 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.http://dx.doi.org/10.1155/2020/8821330
spellingShingle Huandie Yang
Danyang Guo
Leilei Chen
Alexander Suvorov
Qingxin Zhou
Junhua Wang
Xiaofeng Gao
Jinyu Yang
Xiangyan Chen
Application of Nucleic Acid Reference Material for Rapid Detection of Cronobacter sakazakii (Cronobacter spp.) in Flammulina velutipes
Journal of Food Quality
title Application of Nucleic Acid Reference Material for Rapid Detection of Cronobacter sakazakii (Cronobacter spp.) in Flammulina velutipes
title_full Application of Nucleic Acid Reference Material for Rapid Detection of Cronobacter sakazakii (Cronobacter spp.) in Flammulina velutipes
title_fullStr Application of Nucleic Acid Reference Material for Rapid Detection of Cronobacter sakazakii (Cronobacter spp.) in Flammulina velutipes
title_full_unstemmed Application of Nucleic Acid Reference Material for Rapid Detection of Cronobacter sakazakii (Cronobacter spp.) in Flammulina velutipes
title_short Application of Nucleic Acid Reference Material for Rapid Detection of Cronobacter sakazakii (Cronobacter spp.) in Flammulina velutipes
title_sort application of nucleic acid reference material for rapid detection of cronobacter sakazakii cronobacter spp in flammulina velutipes
url http://dx.doi.org/10.1155/2020/8821330
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