InstantLabs Salmonella Species Food Safety Kit
Contents
1. 1 Prepare 3 375 mL of sterile FASTGRO SE Enrichment Broth for each sample to be tested by following the manufacturer s instructions Bring the temperature of the media to approximately 42 C by incubating the prepared media at 42 1 C for 30 60 minutes immediately before use Clean the work area and weighing scale with a 10 bleach solution and wipe until dry Using clean disposable gloves carefully open and place a new Whirl Pak bag onto the laboratory scale Aseptically transfer approximately 750 g of a representative grain sample to the bag using a sterile spatula or scoop Jt is important to try to work quickly to minimize the chances of contaminating the sample Repeat steps 3 and 4 above for each additional sample to be tested Pour 3 375 mL of pre warmed FASTGRO SE Enrichment Broth into each bag and loosely seal bag with built in wire closures f necessary sample size may deviate from 750 g depending on availability of sample as long as the amount of FASTGRO SE is adjusted proportionally Gently hand massage Whirl Pak bag s for 1 2 minutes to mix contents 8 Incubate the Whirl Pak bag s without shaking at 42 C 1 C for 16 20 hours FOR TESTING OF OTHER FOOD SAMPLES 25 g 375 g Sample Collection Homogenization and Enrichment 1 Prepare the recommended volume of sterile FASTGRO SE Enrichment Broth for each sample to be tested by following the manufacturer s instr2. chamber Place the MAC into Hunter and firmly push latch down to lock MAC in place Close the Hunter s door to start the test which will begin in 15 seconds 14 The results will be displayed automatically as Positive Negative or Indeterminate when the cycling is completed TROUBLESHOOTING Problem Suggestion e Positive Control was expired not reconstituted properly stored incorrectly or was not loaded into the MAC correctly Repeat test with fresh reagents No amplification of Positive Control The Hunter calls the positive control Negative or Indeterminate High background in the e Potential contamination Repeat Negative Control test with fresh reagents Hunter calls all samples indeterminate 7 HUNTER SYSTEM SPECIFICATIONS System Components e Hunter device e MAC Assay Cartridges MAC Additional Components e Hunter device User Manual CD e 2 spare fuses e AC line cord Hunter instrument specifications e Dimensions 17 2 cm x 41 2 cm x 22 cm 6 77 in H x 16 22 in W x 8 66 in D e Weight 5 4 Kg 11 9 Ibs e Power usage 90 to 264 VAC at 3 amps e Fuses 2 3 AMP slow blow e Peak block heat rate 4 5 C sec e Peak block cool rate 4 5 C sec e Warm up time 10 minutes e Decibel level less than 40 db at 1 meter Environment e Temperature 10 35 C 50 95 F e Relative humidity up to 90 noncondensing e Locate away from heaters cooling duc
3. 6 0 f ONININ INSTANTLABS NON CLINICAL RESEARCH USE ONLY InstantLabs Salmonella Species Food Safety Kit P N 9034 0600 0003 PRODUCT DESCRIPTION The InstantLabs Medical Diagnostics Corporation InstantLabs Salmonella Species Food Safety Kit contains the reagents necessary for performing qualitative real time PCR on the Hunter Accelerated PCR system see Hunter System Specifications below The kit is intended to be used in an analytical laboratory under standard laboratory conditions KIT STORAGE AND STABILITY Store the InstantLabs Food Safety Kit at 2 8 C The kit is stable through the expiration date printed on the package MATERIALS REQUIRED AND PROVIDED e Multiple Assay Cartridges MAC e Reagent Packs each containing the following o 6 Colored Tubes With Lyophilized PCR Reagents o 4 Sample Extraction Tubes o 1 Tube of Positive Control DNA o 1 Tube of Nuclease Free Water Quantitative PCR complete master mix containing GoTaq Hot Start Polymerase manufactured by Promega Corporation for distribution by BioGX Inc Licensed to Promega under U S Patent Nos 5 338 671 and 5 587 287 and their corresponding foreign patents For Research Use Only Not for any clinical or therapeutic use in humans or animals ADDITIONAL MATERIALS REQUIRED BUT NOT PROVIDED 1 Hunter Accelerated Real Time PCR Instrument 2 Nitrile or Latex Gloves 3 Whirl Pak Filter Bag for Homogenizer Blenders or Other Similar Fil
4. e Table 2 below Table 2 SAMPLE ENRICHMENT TIME Ground Chicken 375 g 10 16 hours Solid Chicken 375 g 10 16 hours Ground Beef 375 g 10 16 hours Beef Trim 375 g 10 16 hours Ground Turkey 375 g 10 16 hours Spinach 25 g 8 16 hours Cabbage 25 g 8 16 hours Lettuce 25 g 8 16 hours Pasteurized Whole Milk 25 g 8 16 hours Cottage Cheese 25 g 8 16 hours Plain Yogurt 25 g 8 16 hours Vanilla Ice Cream 25 g 8 16 hours Salted Butter 25 g 8 16 hours Peanut Butter 25 g 8 16 hours DNA Extraction from Enriched Samples 1 Clean the work area and laboratory pipets with a 10 bleach solution and wipe until dry Firmly tap Sample Extraction Tubes on the laboratory bench to collect contents at bottom of tube Label each tube with the sample to be tested Remove Whirl Pak bag s from incubator being careful not to disturb the settled food debris Carefully open the bag and remove 50 uL of liquid from the top 1 3 cm of the filtered compartment of the Whirl Pak bag without disturbing the settled food debris and place into the correctly labeled Sample Extraction Tube prepared in step 2 above Note that if a layer of fat is present move pipet tip to below the fat layer before removing aliquot Repeat step 4 for each sample to be tested Gently invert Sample Extraction Tube s 3 5 times to ensure sample is mi
5. ly to ensure that lyophilized reagents are reconstituted Additional extracted samples up to a total of 4 can be similarly loaded into the Orange Purple and Yellow tubes if desired Shake the tubes gently to ensure that lyophilized reagents are reconstituted Using a sterile pipet transfer 25 uL of Nuclease Free Water to the blue colored Negative Control tube Shake tube to ensure that lyophilized reagents are reconstituted 6 10 11 12 13 Using a sterile pipet transfer 50 uL of Nuclease Free Water to the tube labeled Positive Control DNA and shake tube vigorously for 15 sec to mix contents Using a new pipet tip transfer 25 uL of this solution to the red colored Positive Control tube Shake tube gently to ensure that lyophilized reagents are reconstituted Briefly spin down the 6 colored tubes in centrifuge for 15 seconds to collect contents at bottom of tube Remove one MAC from the kit Prepare Hunter by entering test and sample information as outlined in the Hunter User Manual Pipette 20u1 of each Sample Control PCR mix solution to the correct port on the MAC by matching the color of the tube to the colors printed on the MAC Insure that the solution flows into the sample chamber and then close the cap on the port To ensure a proper seal press the cap down firmly When all samples have been added to the MAC tap the MAC gently on the countertop to ensure there are no visible bubbles present in the sample
6. ter Bag For 750 g grain samples use 138 oz capacity bags For food samples between 25 g and 375 g use an appropriately sized bag 4 42 1 C Incubator 5 FASTGRO SE Enrichment Broth InstantLabs 6 Sterile Spatula or Scoop Single Use or Re Usable Laboratory Sterilized 7 10 100 uL Pipette with RNase DNase Free Filter Tips 8 Laboratory Balance with a Sensitivity of 1 g 9 Heat Block Set at 95 C 10 Micro Centrifuge 6 000 x g Minimum 11 Stomacher Instrument Optional for most food types but recommended for meat and other samples that cannot be effectively mixed by hand WARNINGS AND PRECAUTIONS 1 a Wear eye protection gloves and laboratory coats when handling samples lysis buffer and kit reagents Avoid contamination of reagents by only using sterile pipette tips Do not use kits beyond the printed expiration date Do not use kits that have been stored above 8 C A negative and a positive control must be included in each test Do not reuse excess reagents from one reagent pack to another dispose of all remaining reagents once all kit MACs have been used Material Safety Data Sheets MSDS are available upon request Treat enriched samples as if they contain pathogenic bacteria Enriched samples and MACs should be disposed of as bio hazardous waste according to local regulations INSTRUCTIONS FOR USE FOR TESTING OF GRAIN SAMPLES 750 g Sample Collection Homogenization and Enrichment
7. ts and keep out of direct sunlight e Protect from all fluids Throughput e Six 6 samples every 2 25 hours run time e Includes pos neg control e 18 samples per 8 hour shift e Supported volumes 20ul Area Requirements e 8 recommended free space around the Hunter system for sample preparation airflow etc TECHNICAL ASSISTANCE Worldwide Support Phone 703 904 4306 Email support instantlabs com Web www instantlabs com 2012 by InstantLabs Medical Diagnostics Corporation Inc All rights reserved
8. uctions See Table 1 below for recommended volumes Bring the temperature of the media to approximately 42 C by incubating the prepared media at 42 1 C for 30 60 minutes immediately before use Table 1 SAMPLE SIZE MEDIA VOLUME 25 g or 25 mL 225 mL 375 g or 375 mL 1 500 mL Media volume and enrichment times of food sample enrichments as a function of desired sample size Any sample size larger than 375 g should be run according to the 750 g grain protocol page 4 Clean the work area and weighing scale with a 10 bleach solution and wipe until dry Using clean disposable gloves carefully open and place a new Whirl Pak bag onto the laboratory scale Aseptically transfer the appropriate amount of a representative food sample to the bag using a sterile spatula or scoop Tt is important to try to work quickly to minimize the chances of contaminating the sample Repeat steps 3 and 4 above for each additional sample to be tested Pour the appropriate volume of pre warmed FASTGRO SE Enrichment Broth see Table 1 above into each bag and loosely seal bag with built in wire closures 7 Gently hand massage Whirl Pak bag s for 1 2 minutes to mix contents Samples containing meats or other difficult to mix materials should be homogenized using a Stomacher machine for 30 60 seconds at maximum speed 8 Incubate the Whirl Pak bag s without shaking at 42 C 1 C for the recommended amount of time se
9. xed with the lysis reagents Incubate the Sample Extraction Tube s at 95 C for 20 minutes in a dry heat block to complete the lysis and then let tube s cool at room temperature for 1 2 minutes Centrifuge tube s in a Micro Centrifuge for 3 minutes at maximum speed to pellet the lysing beads and any food debris The supernatant contains the extracted DNA and is ready to be tested immediately with the InstantLabs Food Safety Kit If the samples will not be tested within 30 minutes transfer 50 uL of supernatant to a new nuclease free microcentrifuge tube and freeze at 20 C for up to 3 months _ _ Avoid Fat Layer If Present Supernatant Contains ONA Avoid Beads and Food Debris Graphic of Sample Extraction Tube Following Centrifugation PCR Setup Overview Each Reagent Pack and Multiple Assay Cartridge MAC can be used to run up to 4 samples plus the required positive and negative controls The tubes in the reagent pack are color coded to match the wells on the MAC and contain convenient ready to use lyophilized reagents Procedure 1 Clean workspace and pipets with a 10 bleach solution and wipe until dry Using clean gloves remove the tubes containing the Lyophilized PCR Reagents from the kit Transfer 25 uL of the extracted sample to be tested to the green colored tube using a sterile pipet being careful to not touch the lyophilized reagents with the pipet tip Shake tube gent
Download Pdf Manuals
Related Search