基于MALDI-TOF MS技术对面膜中污染日勾维多细菌源菌的鉴定分析

doi:10.3969/j.issn.1001-1803.2022.10.011

摘要/Abstract

摘要：

对149批面膜产品进行微生物检验,其中4批菌落总数不符合规定。对不合格样品中分离纯化的菌落用MALDI-TOF MS、16S rDNA测序比对、VITEK 2生化试验3种方法进行鉴定,结果均为日勾维多细菌源菌（Pluralibacter gergoviae）;检出菌与日勾维多细菌源菌标准菌株CICC 24132和一株食品中检出的日勾维多细菌源菌共6株菌在MALDI Biotyper 3及VITEK 2生化反应基础上进行分析分型,经MALDI Biotyper 3对6株菌的蛋白图谱进行聚类分析,来源于面膜中的4株菌亲缘关系比较近;经对比VITEK 2生化试验发现,6株日勾维多细菌源菌生化反应存在差异,其中来源于同一生产厂家的3批面膜检出菌生化反应基本相同。结果表明,MALDI-TOF MS快速鉴定结果可信;经MALDI Biotyper 3聚类分析及生化反应的结果分析,可将4株面膜中分离的日勾维多细菌源菌分成两个亚型,从同一厂家3批不合格样品中检出的污染菌具有高度同源性。

关键词: MALDI-TOF MS技术, 面膜, 日勾维多细菌源菌, 快速鉴定, 同源性分析

Abstract:

A total of 149 batches of facial mask products were tested for microorganism, and 4 batches did not meet the requirements. The bacterial colonies of these 4 batches of facial mask products were purified, and identified by MALDI-TOF MS, 16S rDNA sequencing and Vitek 2 biochemical test. The results show that they are all Pluralibacter gergoviae. In addition, Pluralibacter gergoviae CICC24132 and a food detected Pluralibacter gergoviae are analyzed with these four strains together by MALDI Biotyper 3 and VITEK 2. MALDI Biotyper 3 cluster analysis shows that the protein pattens detected in the four batches of facial mask are basically the same, with their relationship relatively close to each other, but far from the other 2 strains. Biochemical tests of VITEK 2 show that there are differences in the biochemical reactions of 6 strains of Pluralibacter gergoviae, among which the bacteria detected in 3 batches of masks from the same manufacturer are basically the same. The results suggest that the rapid identification results of MALDI-TOF MS are reliable. MALDI Biotyper 3 cluster analysis and biochemical reaction results show that the 4 strains of Pluralibacter gergoviae isolated from facial masks can be divided into two subtypes, and the contaminated bacteria detected from three batches of unqualified samples from the same manufacturer have high homology.

Key words: MALDI-TOF MS, facial mask, Pluralibacter gergoviae, rapid identification, homology analysis

中图分类号:

TQ658

谭慧敏,王涛,朱斌,零文超,樊兰艳. 基于MALDI-TOF MS技术对面膜中污染日勾维多细菌源菌的鉴定分析[J]. 日用化学工业（中英文）, 2022, 52(10): 1107-1112.

Tan Huimin,Wang Tao,Zhu Bin,Ling Wenchao,Fan Lanyan. Identification and analysis of Pluralibacter gergoviae in facial masks based on MALDI-TOF MS[J]. China Surfactant Detergent & Cosmetics, 2022, 52(10): 1107-1112.

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样品编号	不同稀释度平均菌落数			菌落总数/（CFU/g）
样品编号	1∶10	1∶100	1∶1 000	菌落总数/（CFU/g）
HZ095	不可计	74.5	7	7 500
HZ119	不可计	255	27	26 000
HZ120	不可计	不可计	83.5	84 000
HZ121	不可计	263	26.5	26 000

样品编号	VITEK 2鉴定结果	16s rDNA比对结果	MALDI-TOF MS鉴定结果
HZ095	Enterobacter gergoviae（99%）	Pluralibacter gergoviae（>99%）	Pluralibacter gergoviae（2.5）
HZ119	Enterobacter gergoviae（99%）	Pluralibacter gergoviae（>99%）	Pluralibacter gergoviae（2.5）
HZ120	Enterobacter gergoviae（99%）	Pluralibacter gergoviae（>99%）	Pluralibacter gergoviae（2.5）
HZ121	Enterobacter gergoviae（99%）	Pluralibacter gergoviae（>99%）	Pluralibacter gergoviae（2.5）

样品编号	生化反应项目					菌株来源
样品编号	dMAL	CIT	LDC	dCEL	BGAL	菌株来源
HZ095	+	–	+	+	+	化妆品（面膜）
HZ119	–	+	–	–	+	化妆品（面膜）
HZ120	–	+	–	–	+	化妆品（面膜）
HZ121	–	+	–	–	+	化妆品（面膜）
CICC 24132	+	–	+	+	–	临床（小便）
SP80	+	+	–	–	–	食品（卤肉）