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1. 5 Vero toxin VT Hemolytic Uremic Syndrome HUS O157 026 0111 0118 0103 0121 O157 O1572. 100 x L 2 3 D 15 a D
3. 0157 Q MONO 2 D
4. 100L 150 wL 15 6 100 u L 150L
5. 1 30mm 38mm N T RETE LEII L J 1 0157 1 X 10 CFU mL 3 2 0157 3 3 0157 3
6. 1 23 8 2 12 NH O157 20 FARA 1 CE 168 179 2004 2 23 827 834 1996 3 O157 9 7 4 207 199 5 4 0157 9 7 17 T300 2646 3 3 029 847 7825 FAX 029 847 7824 URL http www rdc nipponham co jp 18 11 2 18 2 1 NH 0157
7. Enterobactoer aerogenes Escherichia colt Escherichia colt Enterobactoer aerogenes Escherichia colt Escherichia colt Escherichia colt Escherichia colt Escherichia colt Escherichia colt Escherichia colt Escherichia colt Escherichia colt Escherichia colt Escherichia colt Escherichia colt Enterobactoer aerogenes Enterobactoer aerogenes Escherichia colt Enterobactoer aerogenes Escherichia colt Et A 4 5 4 thit Fy HN HDAIEZ RUNFEOD LIEBE O157 10 EIIELCSABRZCT TO T PS EEE 10 cfu mL 0157 A B Da Coa 0157 10 cfh mL 10 cfu mL 10 cfu m
8. 1 1 2 3 1 2 1 ret eens 5 4 D 0157 nnnm 0 5ug e eee esas 1 Q 0157 0 134 g Q eH 0 25 ug 1 O157 1 0157 1 a F i b d e f 30mm qp C g
9. 1 2 Q RE MDS 200g 300 500cm FES 0 2 0 3mm 25g 3 HEBES D 25g mEC 225mL 1 42 18 24 1 mEC mEC 4mg mL mEC 1 000mL 5mL 20mg L 2 mEC
10. 0157 Kit 0157 2 a p 3 D 121C 20 0 1 1 WV K y VES OCA LOA ETA OBO ECE RESET OB EIIL
11. 38mm a b C d 2 O157 2 Ay 1 0157 1 ll O157 3 O157 2 3 4 0157 4 ZR Eds
12. TSB TSB mTSB TSB CTV mTSB VCC 4 RAIRE D 2 1 2 5mL 121 C 20 1 NH O157 OVO zo E 2 3 4 ED 6
13. 4 1 X104 106CFU mL i Hd 4 Enterobacter cloacae Enterobacter sakazakii Citrobacter freundii Klebsiella pneumoniae Klebsiella oxytoca Serratia liquefaciens Serratia marcescens 8100 2 Citrobacter freundii Salmonella Kumasi 030 0157 18 9 1 18 10 2 NH O157 SE WE S A L FERRE VILE PUPA mEC 7 377 CRE 10 cfu mL 7 109cfu mL
14. Escherichia coli ATCC35150 Ez Escherichia coli ATCC700728 t 1 RCT ER PAR 2 f RIT OBERE RES Escherichia coli NTCC35150 ze PRCA Timi ae pep maT nL ee xp sasse cmm EE l TSB pee p vs 3 f fb t HNDID S Escherichia coli ATCC35150 mEC 3 A B j
15. gt COSMO BIO CO LTD Inspiration for Life Science N ber 2006 2 9 edition Reagent for food testing OYember 2 edition March 2006 1 edition NH IMMUNOCHROMATO O157 Please read this manual carefully before using the kit Instruction Manual Product number 999100000 Introduction Food poisoning by Diarrheagenic Escherichia coli is an infectious food poisoning that is caused by the growth of E coli taken into the intestinal tract though the ingestion of food These Diarrheagenic E coli can be broadly classified into five types Among these Enterohemorrhagic E coli EHEC causes hemorrhagic colitis through the action of the verotoxin that it produces it can also cause hemolytic uremic syndrome and encephalopathy which can be fatal Many cases of mass food poisoning by EHEC have occurred so EHEC is viewed as the most dangerous among Diarrheagenic E coli from the standpoint of food hygiene In addition to E coli O157 other serotypes have been detected such as O26 O111 O118 O103 and O121 In Japan however most of EHEC are reported to be of E coli 0157 This product is a kit for detecting E coli O157 in foods by immunochromatography Tests can be conducted rapidly and simply by means of the kit Product Features 1 The simple one step operation of the kit 2 The test gives rapid results 3 There is no need for special test equipment Kit contents 1 Components 2 Ingredients and Quantity A Test p
16. that there is something abnormal in the development of the sample solution in such cases CIL Notel Ensure that confirmatory tests in accordance with the official test method or other methods are performed on specimens that are tested positive by the kit The proliferated and cultivated samples S a aude used for testing with the kit can be used in confirmatory tests by the official or other test method 1 Sensitivity Test The results of tests conducted in accordance with instructions for the preparation of the sample solution and the operating procedures for testing described in this manual will be positive when the concentration of E coli O157 organisms is more than 1 x 10 CFU mL 2 Repeatability Tests When positive and negative test solutions of E coli O157 were simultaneously tested three times each all positive test solutions exhibited positive results and all negative test solutions showed negative results 3 Minimum Detection Sensitivity The results of testing standard three strains of E coli O157 and four strains of isolated bacteria confirmed that the minimum detection sensitivity is between 1 x 10 and 1 x 10 CFU mL Notel The minimum detection sensitivity of this kit could vary depending on the effects of the components of the test solution 4 Cross reactivity 1 Cross reactivity with the following bacterial strains has not been observed Ennai z9miums Enterobacter arogas O p
17. L 10 cfh mL 10P cfu mL Dye 1 2 0157 3 O157 UNI 5 5 D
18. completed Operating Procedures for Testing 1 NH Immunochromato 0157 Test Procedures 1 Bring the test plates contained in the aluminum pouch to room temperature and remove from the pouch as necessary immediately before use 2 With an oil based marker pen write the name of the test sample or the number of the subject under test on the absorbent pad of the test plate removed from the bag 3 Place the test plate carefully on the flat stand and drop a 100 uL portion of the test solution onto the test sample drop section see the figure on the left Otherwise dispense a 150 uL portion of the test solution into a test tube and attach the test plate to the test tube so that the test sample drop section of the test plate is immersed in the test solution see the figure on the right 4 Allow the test plate to stand undisturbed for 15 minutes and then visually judge the presence or absence of E coli O157 in the solution Drop a 100 uL portion of the test solution Absorbent pad Dispense a 150 uL portion of the test solution Notel Do not remove the test plate from the aluminum pouch until it has returned to room temperature otherwise incorrect test results may be obtained as a result of moisture absorption Note2 Keep test plates that are not in use in a vinyl pouch containing desiccants and store this in an aluminum pouch in a refrigerator Note3 Be careful not to scratch the test sample drop section or expanded section and d
19. e 25g specimen in tomacher bag and homogenize with a stomacher for 1 minute 2 Incubate the specimen in the stomacher bag at 42 C for 18 24 hours Notel When commercial mEC culture medium that does not contain novobiocin is used sterilize the medium by autoclaving and cool it Then sterilize a 4 mg mL solution of novobiocin by filtration and add 5 mL of the solution to 1 L of mEC culture medium to give a final concentration of 20 mg L Note2 Instead of the mEC broth with novobiocin it is possible to use tryptosoya broth TSB or growth and proliferation retardant added TSB mTSB TSB CTV or mTSB VCC selective enrichment broth 4 Sterilization 1 Remove the stomacher bag from the incubator after 18 24 hours Gently mix the contents of the stomacher bag using a side to side motion taking care not to splash it 2 Transfer about 5 mL of the culture solution into a glass test tube by using a sterilized pipette and sterilize the solution in an autoclave for 20 minutes at 121 C 3 Remove the tube from autoclave and allow the tube to cool to room temperature to prepare the sample solution Notel The kit can also detect viable E coli O157 but tests with a sterilized culture solution are recommended to ensure the safety of the operator Note2 Because the remainder of the culture solution might be required for use in confirmatory tests following those conducted with the kit do not sterilize it and retain it until all the tests have been
20. e ATCC4352 Klebsiella oxytoca ATCC8724 Serratia liquefaciens ATCC27592 Serratia marcescens ATCC8100 Serratia marcescens ATCC27143 Serratia marcescens NBRC12648 Serratia ororifia ATCC33077 Salmonella typhimurium ATCC13311 Salmonella enteritidis ATCC13076 Salmonella enteritidis NBRC3313 Salmonella abaetetuba A TCC35640 Salmonella anatum Group E ATCC9270 Salmonella arizonae ATCC13314 Salmonella choleraesuis ATCC10708 Salmonella kunzendorf ATCC12011 Salmonella tallahassee ATCC12002 Salmonella vellore Group B ATCC15611 Vibrio parahaemolyticus NBRC12711T Vibrio vulnificus NBRC15645 Psudomonas aeruginosa ATCC27853 Psudomonas aeruginosa NBRC13275 Aeromonas hydrophila ATCC7966 Staphylococcus aureus NBRC12732 Staphylococcus aureus NBRC3761 MRSA IID1677 Staphylococcus epidermidis NBRC12993 Bacillus cereus ATCC11778 Bacillus cereus NBRC3836 Bacillus subtilis ATCC6633 Bacillus subtilis NBRC3134 Bacillus thuringinensis NBRC13866 375 FARA 7 BIER Listeria monocytogenes ATCC7644 Listeria monocytogenes ATCC15313 Lactobacillus plantarum ATCC8014 Lactococcus lactis subsp actis NBRC12007 3 Rip RE KOKGRARLOXKIGREEORESONE 10 cfu mL
21. e in charge of testing Verify your own operating procedures for the kit and the appropriateness of its use for each particular food Product specifications may be changed without notice 2 Precautions Regarding Risk Prevention 1 2 3 4 Even minute amounts of E coli O157 which the kit is designed to detect could cause infection For this reason and because there is a possibility of infection by microorganisms than other than E coli O157 exercise full precautions in conducting tests by wearing protective gloves and safety glasses Tests should be performed only where appropriate equipment and facilities are available Follow standard microorganism testing procedures under the guidance of responsible supervisors If you accidentally get any sample solution in your eyes or mouth adopt emergency measures such as immediately washing away the solution with tap water and then seek medical attention If you feel unwell after performing a test with the kit obtain immediate treatment from a physician 3 Precautions Regarding Disposal of Waste Materials 1 2 Note that surplus test solutions and used test plates culture media and test samples could carry contagious microorganisms Therefore make sure that waste materials are subject to appropriate sterilization for example by autoclave treatment for 20 minutes at 121 C or immersion of the materials in a sodium chlorite solution for more than 1 hour Discard the k
22. ine appearance position This results in the appearance of a reddish purple line of gold colloid This reddish purple line can be detected by visual 2 3 4 inspection and used to judge the presence or absence of E coli 0 157 in the test solution The excess gold labeled antibodies regardless of the presence or absence of coli O 157 in the test solution travel further through the expanded section and are trapped by the anti goat immunoglobulin rabbit antibody 4 fixed at the control line appearance position where they form a second reddish purple line The presence of this line indicates that the test solution has reached the expanded section www cosmobio co jp COSMO BIO CO LTD Inspiration for Life Science Preparation of the sample Solution The method for preparing a sample solution is described in accordance with the Japanese official testing method 1 Required Equipment and Instruments Stomacher bag preferably with a filter stomacher incubator autoclave culture medium etc 2 Preparation of Test Samples 1 Take a test sample of more than 200g of the food under test In cases where surface contamination is suspected the sample is taken by scraping off 300 500cm of the surface to a thickness of 0 2 0 3mm 2 Chop and mix the whole sample collected Weigh 25g of the sample into the stomacher bag and use this as the test specimen 3 Sample enrichment 1 Add 225 mL of mEC broth with novobiocin to th
23. its test samples and surplus test solutions in strict compliance with your local waste disposal regulations and with full consideration of environmental sanitation Storage Method and Use By Date 1 Storage method Refrigerate at 2 8 C and shade from the light Avoid freezing 2 Use by date 12 months from the date of manufacture Packaging Unit NH Immunochromato O157 20 tests 5 test x 4 References 1 2 3 4 Ministry of Health Labor and Welfare Microorganism Section of Guide to Food Hygiene the Japan Food Hygiene Association pp 168 179 2004 Akemi Kai et al Clinical Diagnosis and Microorganism 23 pp 827 834 1996 Ministry of Health and Welfare Inspection Method for Intestinal Hemorrhagic E coli O157 July 4 1997 Ministry of Health and Welfare Explanation of Inspection Method for Intestinal Hemorrhagic E coli O157 July 17 1997 Manufacturer R amp D Center Nippon Meat Packers Inc 3 3 Midorigahara Tsukuba Ibaraki 300 2646 Japan Phone 81 29 847 7825 Fax 81 29 847 7824 URL http www rdc nipponham co jp RID OF 5 COSMO BIO CO LTD Inspiration for Life Science TOYO 2CHOME KOTO KU TOKYO 135 0016 JAPAN http www cosmobio co jp e mail export cosmobio co jp Phone 81 3 5632 9617 FAX 81 3 5632 9618 1
24. late 5 test x 4 packs A Ant E coli O157 polyclonal antibody rabbit 0 5 ug B Instruction manual 1 sheet B Ant E coh O157 polyclonal antibody labeled with colloidal gold goat 0 13 ug C Anti goat immunoglobulin polyclonal antibody rabbit 0 25 ug Application 1 Detection of E coli O157 in Foods Note 1 This kit is intended to specifically detect E coli O26 and therefore cannot detect the presence or absence of verotoxin prodution Illustration of Test plate and the Principle of assay 1 Illustration of Test plate a Sample solution drop section Be careful not to touch this sectoin with your finger e f g b Reagent containing section c Detecting section Be careful not to scratch this section and touch this section with your finger d Absorbent pad e Measurement items listing position f Testline appearance position Approx 30mm from the _ sample solution drop section 4 4 M g Control line appearance position Approx 38mm from the sample solution drop section 2 Principle of assay W hen a sample solution is dropped onto the test sample drop section of the test plate the gold colloid labeled anti E coli O 157 polycolonal antibody 2 A 1 s in the test sample containing section dissolves and forms complexes with coli 0157 1 These complexes move to the expanded section by capillary attraction and are trapped by the anti co O 157 antibody 3 that is fixed in the test l
25. ml Enerobacter cloacae pgl Enerobacter cloacae awml gyeokoeskgzn sgl Oebeerjemdi SNO S6 U deehpeweue 4523 Klebsellacmtoca Serratia tujen zz Serato marcescens sl amp mwaohrs O mn E 2 There is a possibility that one part of Citrobacter freundii and Salmonella kumasi 030 will show some cross reactivity because they contain the same antigen as F coli 0157 Performance www cosmobio co jp gt COSMO BIO CO LTD Inspiration for Life Science Precautions in Using the Kit 1 Precautions in Handling the Kit 1 2 3 4 5 6 7 Read the instruction manual carefully before use Use the kit in accordance with the test method described in this manual Do not use a kit whose use by date has passed The expiry date is indicated on the label on the aluminum pouch The kit is a reagent designed to detect E coli O157 in food It is not to be used for clinical diagnosis Tests may give false positive results as a result the effects of the ingredients present in the specimen Positive test results from the kit should be confirmed by using the official test method or other procedures Confirm with their manufacturers or distributors that any reagents including culture media required for preparation of test solutions and any instruments that are used are suitable for the purpose This instruction manual is intended as a guideline for thos
26. o not touch them with your hands When handling the test plate make sure that you hold the absorbent pad Note4 Make sure that you use a sterilized pipette or chip to drop or dispense the sample solution Change the pipette or chip for every test solution Note5 Make sure that the 150 uL portion of test solution does not overflow the test plate when dropping it If necessary drop the solution in two or more portions Note6 To prevent infection of the operator it is recommended that testing be performed with the test plate A wrap should be placed under the plate when dropping the test solution www cosmobio co jp gt COSMO BIO CO LTD Inspiration for Life Science 2 Judgment of the Test Results Test line appearance Fono a Puan 1 The test results is judged as positive when a reddish purple line is position 30 mm from position 38 mm from observed at the test line appearance position and at the control line the test sample drop the test sample drop appearance position 15 minutes after the start of the test section section 2 Judge the test results as negative when no reddish purple line is 1 Positive N observed at the test line appearance position but a line is observed at the control line appearance position 3 Retest in cases where no reddish purple line is observed at the control appearance position regardless of the presence or absence of a line at D a 2 Negative the test line appearance position It is likely
27. ut si ee aU 00 n Bn _ pus a 9 A 1 Bl HHKORIRIEXKGRLOZE IME PaclCANL 7c SEPA RO TARE KABA 108cfu mL O157 KH17 77 KH17 3 KH17 125 KH17 72 KH17 6 KH17 12 KH17 225 KH16 8 KH14 144 KH14 209 KH14 263 KH17 159 KH17 201 KH17 28 KH17 178 KH17 306 KH17 13 KH17 162 KH17 79 KH14 247 KH16 131 ag ern re zie O Go O Co RO DS o O S e 2 5 2 HUD BIERELODASZ h INFE lO8cfu mL Esherichia colt ATCC25922 Esherichia colt ATCC1775 Esherichia colt NBRC3972 Esherichia colt NBRC15044 Enterobacter aerogenes ATCC13048 Enterobacter cloacae ATCC13047 Enterobacter sakazakil ATCC8090 Citrobacter freundit ATCC8090 Citrobacter freundit ATCC43864 Klebsiella pneumonia
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