份将检测
机一
些一
Vol.36,
No.18
,2015
食源性致病菌
MALDI-TOF
MS
检测方法的建主与应用
周干渝张延国黄成才
2
谢景丽高青珍文
IJ
肃
1.
(1.中国农业科学院蔬菜花卉研究所,农业部蔬菜品质监督检验圳试中心,北京
100081
;
2.
岛津企业管理(中国)有限公司,分析仪器事业部,北京
100020)
摘
要:建立用基质辅助激光解吸电离飞行时间质谱
(MALDI-TOF
MS)
快速检测并鉴定从鲜食蔬菜中分离到的
5
种
食源性致病菌的方法
f
以标准茵林为模式菌,考察基质溶液自己比、菌株培养条件、溶剂处理方法、点样方法及比例等条
件对鉴定结果的影响,确定最优方法,并考察方法的稳定性、重复性和检测
I'
Lt分别用所建立的方法和
16S
rRNA
基因
测序法对
7
株标准菌株和
93
株分离株进行分析,比较两者鉴定结采的一致性二结果表明,所建立的方法稳定性良好,检
测限为
10
6
-10<)
du/mL
,菌株鉴定结采与基因测序鉴定结果具有较高一致性
(83%
在种水平鉴定一致,
949
毛在属水平鉴
定一致),
MALDI-TOF
MS
法可用于食源性致病菌的准确、快速、低成本、高适量的检测及鉴定,~
关键词
:MALDI-TOF
MS
,食源性致病菌,检测,鉴定
Matrix-assisted laser desorption ionization-time of flight mass
spectrometry for the identification and detection of foodborne pathogens
ZHOU
Qian-yu
l
,
ZHANG
Yan-guo
l
•
HUANG
Cheng-caF. XIE
Jing-l
jI •
GAO
Qing-zhen
l
,LIU
SU
1
'
C
l.
lnstitute of
Vegetables
and
Flowers
,
Chinese
Academv
of
Agricultural
Sciences
,
Supervision
and
Testing
Center
f
Ol
Vegetable
Quality , Ministry of Agriculture , Beijing
100081
,
China;
2.Shimadzu
C
China)
Co.
, Ltd. , Anal ytical
Instruments
Dept
, Beijing
100020
,
China)
Abstract: A method based on matrix - assisted laser desorption ionization - time of flight mass spectrometry
(MALDI-
TOF MS) was developed for rapid discrimination of 5 species of foodborne
pathoge
门
s
which isolated
from fresh vegetables. Standard strains were used
in
the study to investigate factors affecting the identification
results. such as the selection of matrix
solutio
门,
cultivation conditions , solvent treatment method , and the sample
deposition method. After determined the optimal protocol
, the stability and repeatability of the method were
evaluated
, Then bacteria strains separated from fresh vegetables were identified by this protoco
l.
The 16S rRNA
gene sequence analysis for 93 wild strains and 7 standard strains was used as a reference to evaluate the
credibility of
MALDI-
TOF
MS
method. The
MALDI-
TOF
MS
method showed good stability with appropriate
detection limit (
10
6
-10
9
cfu/mL)
for rapid detection of those strains and high consistency with the 16S rRNA
gene sequence analysis (83% consistency at the species level and 94% consistency at the genus level).
It
was
showed that
MALDI-TOF
MS
could be used
as
an
accurate , quick , cost-effective and high throughput method
in
the identification of foodborne pathogens
Key words:
MALDI
一
TOF
MS; foodborne pathogens; detection; identification
中图分类号:
TS207
.4
文献标识码
:A
文章编号
:1002-0306(2015)18-0059-07
doi:
10.
13386/j.issn1002-0306.2015.
18
,
003
食游;但二致病南导致的食品安全问题越来越受到
关注。资料显示,
2006
年到
201
Oi
'f,我因共收到食源
性疾病暴发事件报告
2023
起,累计发病
62920
人,死
亡
967
人,其叶
1
微生物引起的食源性疾病暴发事件数
和
J
忠古者数最多,分另别iJ
l
专叮
ll4
柏
0.0
仰
9%
毛叫叫斗丰和和:可刑
I
门
16
1.
9
归
2
倪%川。
存在
Jr
二食
品介质上的潜在致病菌包括沙门氏
I
岩、大肠杆菌、正在
核细胞增生李斯特菌、金黄色葡萄球菌、蜡样芽胞杆
菌等,这些污染若未得到及时监测手
11
有效控制,贝iJ对
人体存在极大潜在危害
121
0
微生物快边检测技术的应
用将为食源性致病
I
有监测提供重要的技术支撑。
收稿日期
2014-12-05
作者简介:周千;'{j;f(
1991-)
,女,在读硕士研究生,研究方向.食品加工与安全,
E-mail:
q
川
JOU
1991@163.
C'
olll
1
通讯作者:)Ì
I]
肃(
1955
一),男,硕士,研究员,研究方向-农产品质量和安全检测技术.
E-Ill<:li!:
liu:.m@)lllui
l.
c
Cl
HS.
Il
C
t.
C
Il
基金项目,
2014
年国
t
家蔬菜产品质安安全风险坪估项目
(CJFP20
1400
1 )
(EEEEE
二
59