農研機構研究報告 食品研究部門 第1号
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1 Bull. NARO. Food. ResNo., 研究ノート 3M TM 病原菌自動検出システム改良サルモネラ検出キット MDA2 SAL の評価 Fia Noviyanti Evaluation of 3M TM Molecular Detection System MDA2 SAL for the detection of Salmonella from food samples Susumu Kawasaki* 1, Mie Saito 1, Fia Noviyanti 2, Takehisa Harada 3, Shoko Saito 4, and Takatoshi Moriyama 4 1 Food Research Institute, National Agriculture and Food Research Organization, Kannondai, Tsukuba, Ibaraki ) 2 Tsukuba University, Tennoudai, Tsukuba, Ibaraki ) 3 Maruha Nichiro Corporations, 16-2 Wadai, Tsukuba, Ibaraki , Japan) 4 3M Health Care Limited, 33-1, Tamagawadai 2-chome Setagaya-ku, Tokyo ) Abstract Molecular detection Assay2 (MDA2) SAL, the isothermal amplification with bioluminescent assay has recently been available as modified version of previous Salmonella detection kit (MDA1). The performance of MDA2 SAL was compared to MDA1, traditional PCR, and conventional culture (NIHSJ-01-ST4) assay. The MDA2 assay was 100% specific among 13 strains (6 Salmonella and 7 non-salmonella). The detection limits were 10 3 to 10 4 CFU per ml in spiked food homogenate samples without enrichment step. The excellent agreement was obtained for the results of MDA2 SAL kit and conventional culture method from artificially contaminated food samples ( CFU/25g) with enrichment step. These results suggest that the MDS assay is a reliable and useful method for rapid screening of Salmonella contamination. Key words: 3M Molecular detection system, Salmonella, rapid detection, foodborne pathogens Corresponding [email protected]
2 HACCP S. Enteritidis HACCP Codex HACCP M Molecular Detection System MDS Molecular Detection Assay kitmda Listeria monocytogenes MDA Molecular Detection Assay MDA MDA FDA/BAMMDA MDA MDAMDAPCR 実験材料および方法 1. 供試菌および供試食材と接種菌数の確認 S. TyphimuriumO: FSDS. ThompsonO: FSDS. Enteritidis O: FSDS. SenftenbergO,,: FSD S. NewportO: FSDS. WeltevredeneO,: FSD Citrobacter diversusfsdc. freundii FSDEscherichia colifsdenterobacter cloacaefsdcronobacter sakazakiifsd Psuedmonas aeruginosanbrcproteus vulgaris NBRCTrypticase Soy BrothTSB, Difco LTripticase Soy Agar TSA; DifcoTSA
3 2. 培養液および供試食材を含む培養液からのサルモネラ検出限界確認試験 MDAMDA CFU/mLMDA MDA g mlbuffered peptone waterbpw; ISO Pro-media SHM; ELMEX ml ml CFU/mL MDAMDA 3. 特異性試験 MDAMDA CFU/mL CFU/mL LMDAMDA 4. 前培養工程を含めた検出感度試験 g CFU CFU NIHSJ--ST LMDAMDA PCR ml PCR PCR TAPathogenic Bacterial Multiplex PCR Detection SystemDNADNA Extraction Kit TAMultiplex PCR Pathogenic Bacterial Multiplex PCR Detection Kit TA Takara DNA PCR PCR PCR l.sigma UV -bp NIHSJ--ST 5.MDA2およびMDA1による核酸抽出法および遺伝子増幅反応による検出 MDS M Molecular Detection Assay SALM M Molecular Detection InstrumentM L MDAM Lysis tubelysis tube L M Molecular Detection Instrument / MDAMDA 実験結果および考察 MDAMDA CFU/mL MDAMDA MDAMDA MDA MDA
4 表 1. 各サルモネラ菌液および供試食材を含む菌液からの検出限界確認試験結果. Strain serotype Log CFU/mL MDA MDA MDA MDA MDA MDA MDA MDA MDA MDA MDA MDA MDA MDA S. Thompson O S. Senftenberg O S. Typhimurium O S. Enteritidis O,, S. Newport O S. Weltevreden O, mllog/ml 表 2.MDA2, MDA1 キットによる特異性試験結果 Strain MDA MDA Salmonella Typhimurium Thompson Enteritidis Senftenberg Newport Weltevreden Citrobacte diversus freundii Escherichia coli Enterobacter cloacae Cronobacter sakazakii Psuedmonas aeruginosa Proteus vulgaris 表 3. 供試食材に S. Typhimurium を 10 0 CFU/25g 接種した検体からの各手法による検出結果 MDA MDA PCR MDA BPW CFU/mL CFU/mL LL L MDAMDA. CFU/mLLog. CFU/mL MDAMDA OO OOOOO Citrobacter BPWMDA MDA PCR MDAMDA MDA MDA L L MDA MDA
5 MDA 参考文献 FDA: Bacteriological Analytical Manual, Capture (). MTM Molecular Detection System Listeria monocytogenes 80- Bird, P., Flannery, J., Crowley, E., Agin, J. R., Goins, D., and Monteroso, L., Evaluation of MTM Molecular Detection Assay (MDA)-Salmonella for the detection of Salmonella spp. in select foods and environmental surfaces: Collaborative study, First action.. J. AOAC Int. 99, -.(). Kawasaki, S,. Horikoshi, N., Takeshita, K., Sameshima, T., and Kawamoto, S., Multiplex PCR for simultaneous detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O:H in meat samples. J. Food Prot. 68, -. (). 要約 M Molecular detection Assay SALMDA SAL MDA SAL MDA MDAPCR MDA CFU/mL MDA MDAMDA NIHSJ--STPCR MDA
緒言 Listeria monocytogenes ready-to-eat L. monocytogenes ISO - FDA/BAM L. monocytogenes Real-Time PCR DNA PCR SYBR Green ds-dna PCR LAMP Molecular Dete
Rep. Nat l Food Res. Inst No 報 文 3M TM Molecular Detection System を用いた Listeria monocytogenes の 簡易迅速遺伝子検査法の評価 Evaluation of 3M Molecular Detection System for detecting Listeria monocytogenes from food
320 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /., No.1, -,* -,/ (,**1) 8 * ** *** * ** *** E#ect of Superheated Steam Treatment on the Preservation an
320 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /., No.1, -,* -,/ (,**1) 8 * ** *** * ** *** E#ect of Superheated Steam Treatment on the Preservation and Sensory Characteristics of Buckwheat Noodles Kazuhiro
Staphylococcus sp. K.pneumoniae P.mirabilis C.freundii E. cloacae Serratia sp. P. aeruginosa ml, Enterococcus avium >100ƒÊg/ml
CHEMOTHERAPY SEPT. 1992 cefoperazone ceftazidime (CAZ), imipenem (IPM) Staphylococcus sp., Enterococcus (CPZ), faecalis, Escherichia coli, Klebsiella pneumoniae, Citrobacter freundii, Enterobacter cloacae,
はじめに
LAMP 法を用いたウェルシュ菌エンテロトキシン遺伝子検出の検討 佐藤秀美 Development of detection of Clostridium perfringens enterotoxin gene using the LAMP assey Hidemi Sato はじめにウェルシュ菌による食中毒は, 大量に調理された食品を原因とすることが多く, しばしば大規模になるため, 原因物質の迅速な検出と判断が求められている.
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