市售婴幼儿配方奶粉和婴幼儿米粉中梭状芽胞杆菌的分离和综合鉴定分析
作者:
作者单位:

北京市疾病预防控制中心营养与食品卫生所,北京市预防医学研究中心,食物中毒诊断溯源技术 北京市重点实验室,北京 100013

作者简介:

陆峥 女 副主任技师 研究方向为食品微生物 E-mail:luzheng0715@163.com

通讯作者:

张晓媛 女 研究员 研究方向为食源性致病菌的分子流行病学 E-mail:zhangxiaoai_0922@163.com

中图分类号:

R155

基金项目:

首都高层次公共卫生人才建设项目(学科骨干-01-027)


Isolation and comprehensive identification of Clostridium sp. in commercially available infant formula milk powder and infant formula rice flour
Author:
Affiliation:

Beijing Center for Disease Prevention and Control, Beijing Center for Disease Preventive Medical Research, Beijing Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning, Beijing Center for Disease Prevention and Control, Beijing 100013, China

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    摘要:

    目的 对57件市售婴幼儿配方奶粉、50件婴幼儿米粉进行梭状芽胞菌分离鉴定分析及毒素基因检测,获得梭状芽胞杆菌的污染水平数据,并评估各鉴定方法。方法 通过分离菌株的生长特性、革兰染色、普通显微镜下形态特征等表型特征,应用微生物飞行时间质谱(MALDI-TOF-MS)、16S rRNA基因测序技术、全基因组测序技术对分离菌株进行综合鉴定,通过PCR对分离的梭状芽胞杆菌进行肉毒毒素基因检测,对肉毒毒素基因PCR阳性片段进行测序和比对分析。结果 57份市售婴儿配方奶粉中有26份样品中检出38株梭状芽胞杆菌,其中9份样品中同时检出超过两种细菌。50份市售婴幼儿米粉中有5份样品检出5株梭状芽胞杆菌。奶粉中分离的一株楔形梭菌E型肉毒梭菌毒素基因PCR扩增阳性,扩增片段测序比对结果显示这段序列并不是E型肉毒梭菌毒素基因,全基因组测序结果也证实。43株梭状芽胞杆菌均未检出肉毒梭菌的毒素基因。结论 梭状芽胞杆菌鉴定需要多种方法的综合分析。市售婴幼儿配方奶粉、婴幼儿米粉中存在梭状芽胞杆菌的污染,应加强婴幼儿配方食品中重要梭状芽胞杆菌的监测,为风险评估提供数据支持。

    Abstract:

    Objective Clostridium sp. isolation, identification and analysis and toxin gene detection were carried out on 57 pieces of commercially available infant formula milk powder and 50 pieces of commercially available infant rice noodles sold in Beijing. The data of Clostridium sp. contamination level was obtained, and the identification methods were evaluated.Methods The isolates were comprehensively identified by their growth characteristics, Gram staining, morphological characteristics under ordinary microscopy, combined with the results of mass spectrometry identification, 16S rRNA gene sequencing and whole-genome sequencing (WGS). The botulinum toxin genes of isolated Clostridium sp. were detected by PCR. Then, sequencing and comparative analysis of PCR positive fragments were carried out.Results Thirty-eight Clostridium sp. in 26 samples of the 57 commercially available infant formula milk powder samples were detected. More than two types of isolates were detected in 9 samples. Five Clostridium sp. were detected in 5 samples of 50 commercially available infant rice flour. PCR amplification of the Clostridium botulinum type E toxin gene of a Clostridium sphenoides was positive, the amplified fragment sequencing comparison results showed that this sequence was not an E-type Clostridium botulinum toxin gene, and the WGS results also confirmed it. There were no toxin genes of Clostridium botulinum in all the 43 isolates.Conclusion Identification of Clostridium sp. requires comprehensive analysis by multiple methods. The contamination of Clostridium sp. exists in commercial infant formula milk powder and infant formula rice flour. It is necessary to strengthen the monitoring of important Clostridium sp. in infant formula food to provide data support for risk assessment.

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陆峥,刘玉竹,张鹏航,张诣,陈倩,张晓媛.市售婴幼儿配方奶粉和婴幼儿米粉中梭状芽胞杆菌的分离和综合鉴定分析[J].中国食品卫生杂志,2024,36(4):433-439.

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  • 收稿日期:2023-04-04
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  • 在线发布日期: 2024-07-04
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