多重核酸检测系统联合MALDI-TOF MS在产气荚膜梭菌鉴定中的应用
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作者:
作者单位:

1.张家港市疾病预防控制中心,江苏 张家港 215600;2.江阴市食品安全检测中心,江苏 江阴 214400;3.上海理工大学健康科学与工程学院,上海 200093;4.张家港海关综合技术中心,江苏 张家港 215600

作者简介:

施菊萍 女 副主任技师 研究方向为微生物检验 E-mail:1006824137@qq.com

通讯作者:

王翔 男 副教授 研究方向为食品微生物 E-mail:xiang.wang@usst.edu.cn

中图分类号:

R155

基金项目:

江苏省卫生健康委科研课题(Ym2023077);苏州市预防医学科研课题(SZYZ202409);苏州市“科教兴卫”青年科技项目(KJXW2023066)


Application of multiple nucleic acid detection system combined with MALDI-TOF MS in the identification of Clostridium perfringens
Author:
Affiliation:

1.Zhangjiagang Centre for Disease Control and Prevention, Jiangsu Zhangjiagang 215600, China;2.Jiangyin Food Safety Testing Centre, Jiangsu Jiangyin 214400, China;3.School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China;4.Zhangjiagang Customs Integrated Technical Centre, Jiangsu Zhangjiagang 215600, China

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    摘要:

    目的 建立多重核酸检测系统联合基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)对产气荚膜梭菌的快速筛查鉴定方法,为食源性致病菌检测提供新策略。方法 采集张家港市一起食物中毒事件的样本,分别采用多重核酸检测系统和MALDI-TOF MS快速筛查、传统生化和MALDI-TOF MS进行致病菌鉴定,并以荧光定量PCR结合16S rRNA测序验证鉴定结果的准确性。结果 经多重核酸检测系统和MALDI-TOF MS快速筛查,初步提示了样本中存在产气荚膜梭菌。采用国标GB 4789.13—2012方法检测,在与食物中毒相关的122份样本中检出了31株产气荚膜梭菌。此外,对7株生化试验结果有偏差的菌株进行了荧光定量PCR和16S rRNA基因测序验证,最终确认此7株菌株同样为产气荚膜梭菌。结论 多重核酸检测系统具有快速提示和排除常规检测项目的优势,MALDI-TOF MS操作简便、快速、鉴定结果准确性高,两种技术结合使用,可快速、准确地查找确认致病因子,为食源性致病菌鉴定提供了新的有效手段。

    Abstract:

    Objective To develop a rapid method for the screening and identification of foodborne (C. perfringens) isolates using multiple nucleic acid detection system combined with matrix assisted laser desorpt ionionization time of flight mass spectrometry (MALDI-TOF MS), and to provide a new strategy for the identification of foodborne pathogens.Methods Samples from a food poisoning incident in Zhangjiagang City were collected for rapid screening with the multiplex nucleic acid detection system and MALDI-TOF MS, detection with traditional biochemical and MALDI-TOF MS, the accuracy of the results was verified by Real-time PCR combined with 16S rRNA sequencing.Results After initial screening using a multiplex nucleic acid detection system and MALDI-TOF MS, preliminary evidence indicated the samples had C. perfringens. Following GB 4789.13—2012 testing protocol, 31 strains of C. perfringens were identified in 122 samples associated with food poisoning incidents. Furthermore, fluorescence quantitative PCR and 16S rRNA gene sequencing were employed to verify 7 strains that showed inconsistent biochemical test results. These additional analyses ultimately confirmed that these 7 strains were indeed C. perfringens.Conclusion The multiple nucleic acid detection system has the advantages of prompt detection and elimination of routine detection items, and the MALDI-TOF MS is easy to operate, rapid, has a high accuracy of detection results. The combination of the two technologies can quickly and accurately screen and confirm pathogenic factors, providing a new effective method for the detection of foodborne pathogens.

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施菊萍,王亚平,刘芳,徐兰,钱惠芬,樊飞,王翔,邵景东.多重核酸检测系统联合MALDI-TOF MS在产气荚膜梭菌鉴定中的应用[J].中国食品卫生杂志,2025,37(2):131-137.

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  • 收稿日期:2024-05-31
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  • 在线发布日期: 2025-06-05
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