7 6 Vol. 7 No. 6 2016 6 Journal of Food Safety and Quality Jun., 2016 1 O157 阚式绂 1, 黄裕 1*, 王明明 (1., 518005; 2., 518083) 2 摘要 : 目的 sznjo23 方法 sznjo23, 结果 CT-SMAC, CHROMagar O157 VITEK 2 Compact, 96% O157,, 16S rdna, (ATCC29903), 99.26% 结论 sznjo23 16S rdna, 关键词 : ; ; ; 16S rdna Identification of a strain of Shigella with biochemical features similar to Escherichia coli O157 KAN Shi-Fu 1, HUANG Yu 1*, WANG Ming-Ming 2 (1. Shenzhen Inspection and Testing Centre for Quality and Safety of Farm Products, Shenzhen 518005, China; 2. BGI-Shenzhen, Shenzhen 518083, China) ABSTRACT: Objective To analyze and identify a bacterium of sznjo23. Methods A strain of sznjo23 was isolated from pig s liver, and the colonial morphology, biochemical features, and molecular biological characteristics of the strain were analyzed. Results Its colonies features on CT-SMAC were rounded, smooth, small, and colorless, while the strain became purple and circular on CHROMagar O157. Biochemical results showed that the strain had 96% possibility to be E. coli O157 by VITEK 2 Compact. Serology reaction was cloudy, and it didn t appear any agglutination. The sequence analysis result of 16S rdna showed that the strain was closed to Shigella (ATCC29903) from evolutionary distance with the similarity of 99.26%. Conclusion The strain of sznjo23 was Shigella. When the results of biochemical tests and serological test were inconsistent, 16S rdna sequence analysis can be used in order to ensure the accuracy of the results. KEY WORDS: Shigella; biochemical tests; serological test;16s rdna sequence 1 引言, [1],,, * 通讯作者 :,, E-mail: 910241198@qq.com *Corresponding author: HUANG Yu, Senior Veterinarian, Shenzhen Inspection and Testing Centre for Quality and Safety of Farm Products, Shenzhen 518005, China. E-mail: 910241198@qq.com
6, : 1 O157 2259 [2],,,, [1] (EHEC), EHEC 50, O157:H7 [3] O157:H7 1982, [4], O157:H7, 1996, O157 [5] 1999 O157:H7 [6],, [7],, O157:H7 [8,9] O157:H7 [10],, sznjo23, 16S rdna,, ATCC29903 16S rdna O157:H7,, 16S rdna,, 2 材料与方法 2.1 实验材料 : IPP110 ( Memmert ); HEV-50 ( HIRAYAMA ); VITEK 2 Compact( ) : EC (CT-SMAC) CHROMagar O157 (TSI); VITEK 2 GN ; O157 S&A : 2015 : O157 NCTC12900 CMCC51572 2.2 实验方法 2.2.1 取样与增菌 GB/T 4789.36-2008 O157: H7/NM : 25 g, 225 ml mec+n, 1 min, 37 18~24 h O157 NCTC12900 CMCC51572 2.2.2 分离培养 O157 NCTC12900 CT-SMAC CHROMagar O157, 37 18~24 h 2.2.3 生化试验及血清学试验, TSI TSI, O157 VITEK 2 Compact GN 2.2.4 16S rdna 序列测定 16S rdna 3 结果与分析 3.1 分离培养结果 1A B, CT-SMAC CHROMagar O157, sznjo23 1C D, O157 NCTC12900 CT-SMAC, CHROMagar O157 3.2 生化及血清学试验结果 sznjo23 NCTC12900 CMCC51572, sznjo23 96% O157 47, L- α- O/129 L- ELLMAN 7 NCTC12900 ; β-d-l- L- L- 9 CMCC51572 NCTC12900 97% O157 CMCC51572 99% 1 sznjo23,, 3.3 16S rdna 序列测定结果 NCBI, BLAST, MEGA6, sznjo23 16S rdna,
2260 7 Fig. 1 1 sznjo23 Colonial morphology of the bacterium sznjo23 表 1 分离菌株 sznjo23 和标准菌株生化结果 Table 1 Biochemical characteristics of sznjo23, NCTC12900 and CMCC51572 sznjo23 NCTC12900 CMCC51572 TSI 续表 1 sznjo23 NCTC12900 CMCC51572 TSI D- - - - - - - - - - D- - - - - - - - - - L- - - - D- - - - β- + + - H 2 S - - - β-n- - - - pna - - - Y- - - - / + + + β- - - - D- + + - β- - - - β- pna - - - L- - + + - - - - - - + + + - - - ( ) - - - - - - 5- - - - - L- + + - α- - - - + + - N--β- - - - α- - + - - + - - + + + + - + + - - - - COURMARATE + + + β- - - - O/129 + - + - - - - - L- - + - ELLMAN - + - L- - - + : +; -
6, : 1 O157 2261 Fig. 2 2 sznjo23 16S rdna Phylogenic tree of the bacterium sznjo23 based on 16S rdna sequence (neighbor-joining) ( 2), sznjo23 16S rdna (ATCC29903), 99.26% GENBANK, 99% [11-12], 4 讨论与结论 O157 GB/T 4789.36-2008 O157: H7/NM O157:H7,, FDA [13], [14],, 16S rrna 14 [15] ; 16S rdna [16],,,,,,,, sznjo23 96% O157,, sznjo23, 16S rdna, NCBI BLAST, MEGA6, 16S rdna sznjo23 (ATCC29903), 99.26% 99%,,,,, [15] 16S rdna,,, 16S rdna, [17-18],,,, 16S rdna, 参考文献 [1]. [M]. :, 2007. Hirsh. Veterinary Microbiology [M]. Beijing: Science Press, 2007. [2],. O157 [J]., 2013, 29(7):612 614. Zhang ZH, Xi DR. Isolation and identification of bacteria cross-reacting with Shigella and E.coli O157 anti-serums[j]. J Prev Med Inf, 2013, 29(7):612 614. [3]. [M]. :, 2007. Zhou TY. Diagnosis and illustration of clinical microbiology [M]. Shanghai: Shanghai Scientific Technical Publishers, 2007. [4],,. O157 PCR [J]., 2009, 24(6): 438 439. Lu S, Bai L, Xiong YW. A PCR method of identification for Citrobacter freundii agglutinating with O157 serum [J]. Dis Surv, 2009, 24(6):
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