7 7 Vol. 7 No. 7 2016 7 Journal of Food Safety and Quality Jul., 2016 李琼琼, 杨 * 燕, 范一灵, 杨美成 (, 201203) 摘要 : 目的 方法 (1~100 CFU/10 g) (0~10 6 CFU/10 g), GB4789.4-2010, (VIDAS30) (BAX Q7 System), 结果, VIDAS BAX ; 3, XLD BS CAS, CAS, ; BPW 8 h 18 h 结论, 2, VIDAS BAX 关键词 : ; ; Evaluation on the interference of Pseudomonas aeruginosa to Salmonella detection in food samples LI Qiong-Qiong, YANG Yan, FAN Yi-Ling, YANG Mei-Cheng * (Shanghai Institute for Food and Drug Control, Shanghai 201203, China) ABSTRACT: Objective To evaluate the interference of Pseudomonas aeruginosa to Salmonella detection in food samples. Methods The sterile milk powder was artificially contaminated with different concentrations of Salmonella (1~100 CFU/10 g) and Pseudomonas aeruginosa (0~10 6 CFU/10 g). Then those samples were detected according to the GB 4789.4-2010. Meanwhile, VIDAS and BAX System were used to evaluate the interference of Pseudomonas aeruginosa to Salmonella detection. Results Under the interference of different concentrations of Pseudomonas aeruginosa, all Salmonella positive samples could be detected accurately by VIDAS and BAX System. Three kinds of selective culture plates had difference during the separation of Salmonella and Pseudomonas aeruginosa, XLD plate showed better separation ability than BS plate and CHROMagar CAS plate, and Salmonella and Pseudomonas aeruginosa showed similar colony morphology in 基金项目 : (SKT-2015-02-2) Fund: Supported by Shanghai Institute for Food and Drug Control Key Project (SKT-2015-02-2) * 通讯作者 :,, E-mail: yangmeicheng@vip.sina.com *Corresponding author: YANG Mei-Cheng, Chief Pharmacists, Shanghai Institute for Food and Drug Control, No.1500, Zhangheng Road, Pudong District, Shanghai 201203, China. E-mail: yangmeicheng@vip.sina.com
2646 7 CHROMagar CAS plate, which directly affected the detection result of salmonella. The separation ability of pre-enrichment 8 h in BPW was better than pre-enrichment 18 h for salmonella detection. Conclusion It indicated that Pseudomonas aeruginosa showed interference effect during the separation of Salmonella from selective culture plates, using more than 2 kinds of selective culture plates simultaneously could improve the detection efficiency and accuracy. VIDAS and BAX System could be used as the reliable reference of Salmonella detection in food samples. KEY WORDS: Salmonella; Pseudomonas aeruginosa; interference 1 引言 (Salmonella spp.),, [1], 2 [2,3] (WHO), 70%~80% [4],,, [5],,, [6], [7,8],,, 2014 (Food Analysis Performance Assessment Scheme, FAPAS) M195d071,, 74%, 1 (http://fapas.com/),,, 2 材料及方法 2.1 试验菌株 (CMCC(B) 50094) (CMCC(B) 10104) (CMCC), (VITEK2 COMPACT) 2.2 培养基及试剂 (BPW) (SC) (XLD)/ (TSI) ; (TTB) ; (BS) (CHROMagar)(CAS) ; VIDAS Salmonella ; BAX System Real-Time PCR Assay Salmonella GB 4789.28-2013 2.3 主要仪器 NuAire LABGARD SANYO IMR-254 SVW103 (); (VIDAS30, ); (VITEKT 2 COMPACT, ); (BAX Q7 System, ) 2.4 试验方法 2.4.1 菌液制备 (CMCC(B) 50094) (CMCC(B) 10104), (36±1) 18 h, 10, 2.4.2 样品制备, 10 g, 1,,, 3, 27
7, : 2647 表 1 人工污染奶粉样品及沙门氏菌和铜绿假单胞菌的接菌量 (n=3) Table 1 Tested milk powder samples and artificial contamination concentration of Salmonella and Pseudomonas aeruginosa (n=3) a (CFU/10 g) A1 A(B) b A(B) b 0(0) A2 1(0.6) 10 3 (620) A3 10 6 (620000) B1 0(0) B2 10(7.2) 10 3 (620) B3 10 6 (620000) C1 0(0) C2 100(66.5) 10 3 (620) C3 10 6 (620000) : a: 3 ; b: A, B 2.4.3 前增菌和选择性增菌,, BPW, BPW 2,, BPW 100 ml (36±1) 4 8 18 h, 1 ml, 10 ml TTB(42 18~24 h) 10 ml SC (36 18~24 h), 2 2.4.4 沙门氏菌快速筛查 (VIDAS 30) (BAX Q7 System),, 27 4 8 18 h TTB SC 2.4.5 选择性平板分离 27 8 h 18 h TTB SC, BS (36 48 h) XLD (36 24 h), 2.4.6 生化鉴定 GB 4789.4-2010, BS XLD, 2 ;,, 5, 2.4.7 沙门氏菌阳性结果判断标准 VITEK2 Compact 1, 3 结果与分析 3.1 VIDAS 和 BAX 的沙门氏菌快筛结果 3.1.1 不同沙门氏菌和铜绿假单胞菌污染水平 VIDAS BAX 27, 2 1~100 CFU/10 g, 10~10 6, GB 4789.4-2010 (8 h 18 h), 2, A, 3/3, (1 CFU)A 0.6 CFU/10 g Table 2 表 2 VIDAS 和 Bax 沙门氏菌快筛结果 Test screening results of Salmonella by VIDAS and BAX System 4 h 8 h 18 h TTB SC TTB SC TTB SC A1 2/3 a 1/3 2/3 a 2/3 2/3 2/3 A2 1/3 2/3 2/3 2/3 2/3 2/3 A3 0/3 0/3 2/3 2/3 2/3 2/3 B1 2/3; 3/3 b 3/3 3/3 b 3/3 3/3 3/3 B2 2/3; 3/3 b 3/3 3/3 3/3 3/3 3/3 B3 2/3 3/3 3/3 3/3 3/3 3/3 C1 2/3; 3/3 b 3/3 3/3 3/3 3/3 3/3 C2 1/3; 2/3 b 3/3 3/3 3/3 3/3 3/3 C3 1/3; 2/3 b 3/3 3/3 3/3 3/3 3/3 : a ; b VIDAS BAX, VIDAS BAX
2648 7 3.1.2 不同前增菌时间 (4 8 18 h) 2 4 h 8 h 18 h,, 8 h 18 h, 4 h,, A3, 4 h, TTB SC, A1 A2 B3,, 3.2 沙门氏菌传统培养方法检测结果, 27, 3 A 3, 24, VIDAS BAX, BS XLD VITEC 2 Compact, XLD BS 8 h 18 h, ( BS A2 A3 C2 C3 XLD A3 ),, XLD BS CAS ( 3 ), CAS 5, 8 h 18 h, 22.2%(12/54) 14.8%(8/54) CAS ( 4),, CAS 表 3 铜绿假单胞菌干扰下沙门氏菌 3 种选择性平板分离与 VITEK 生化鉴定结果 Table 3 Salmonella detection results of three selective culture media under Pseudomonas aeruginosa interference TTB SC BS XLD CAS BS XLD CAS a b A1 2/3 c 2/2 d 2/3 2/2 2/3 2/2 2/3 2/2 2/3 2/2 2/3 2/2 A2 2/3 2/2 2/3 2/2 3/3 2/3 2/3 2/2 2/3 2/2 3/3 0/3 A3 2/3 2/2 2/3 2/2 3/3 2/3 2/3 2/2 2/3 2/2 3/3 2/3 B1 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 8 h B2 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 1/3 B3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 0/3 C1 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 C2 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 1/3 C3 3/3 3/3 3/3 3/3 3/3 2/3 3/3 3/3 3/3 3/3 3/3 1/3 A1 2/3 2/2 2/3 2/2 2/3 2/2 2/3 2/2 2/3 2/2 2/3 2/2 A2 1/3 1/1 2/3 2/2 3/3 2/3 1/3 1/1 2/3 2/2 3/3 1/3 A3 2/3 2/2 2/3 2/2 3/3 2/3 0/3 0/0 1/3 1/1 3/3 1/3 B1 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 18 h B2 3/3 3/3 3/3 3/3 3/3 2/3 2/3 2/2 3/3 3/3 3/3 3/3 B3 3/3 3/3 3/3 3/3 3/3 2/3 2/3 2/2 3/3 3/3 3/3 2/3 C1 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 C2 2/3 2/2 3/3 3/3 3/3 3/3 2/3 2/2 3/3 3/3 3/3 1/3 C3 2/3 2/2 3/3 3/3 3/3 3/3 2/3 2/2 3/3 3/3 3/3 3/3 : a ; b VITEK2 Compact
7, : 2649 表 4 铜绿假单胞菌干扰下沙门氏菌 CAS 平板选择性分离结果 Table 4 Salmonella separation results of CHROMagar CAS medium under Pseudomonas aeruginosa interference 8 h 18 h TTB SC TTB SC A1 5/5 a ; 5/5; / 5/5; 5/5; / 5/5; 5/5; / 5/5; 5/5; / A2 5/5; 5/5; / 0/5; 0/5; / 1/5; 4/5; / 0/5; 3/5; / A3 5/5; /; 5/5 2/5; /; 3/5 1/5; /; 1/5 1/5; /; 0/5 B1 5/5; 5/5; 5/5 5/5; 5/5; 5/5 5/5; 5/5; 5/5 5/5; 5/5; 5/5 B2 5/5; 5/5; 5/5 2/5; 0/5; 0/5 0/5; 3/5; 5/5 3/5; 3/5; 5/5 B3 5/5; 5/5; 5/5 0/5; 0/5; 0/5 2/5; 0/5; 3/5 0/5; 3/5; 4/5 C1 5/5; 5/5; 5/5 5/5; 5/5; 5/5 5/5; 5/5; 5/5 5/5; 5/5; 5/5 C2 5/5; 5/5; 5/5 0/5; 2/5; 0/5 5/5; 1/5; 4/5 0/5; 3/5; 0/5 C3 4/5; 0/5; 5/5 0/5; 15; 0/5 4/5; 0/5; 4/5 1/5; 2/5; 5/5 : a 3 ; /, VIDAS BAX 3.3 沙门氏菌和铜绿假单胞菌在 3 种选择性平板上的菌落形态比较 3, 1 5 XLD,, A3-18h -SC-XLD 1 ( 3), XLD BS,, BS, 48 h,,,, ( 4 ) 4 讨论,, 10~10 6, GB4789.4-2010, TTB SC 2, BS-XLD BS-,, [9],, BPW 8 h BPW 18 h BPW,,,,,, 图 1 沙门氏菌和铜绿假单胞菌在 BS 平板 XLD 平板以及沙显平板上的菌落形态 Fig. 1 Colonial morphology of Salmonella and Pseudomonas aeruginosa c in BS, XLD, CHROMagar CAS plate.
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