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LiliF® VTEC Real-time PCR Kit (ver.2.0)

Cat.No Capacity Inquire
IP24739 50 Tests Inquire
PRODUCT INFORMATION

 

Background Information
Food microbiological test kit applied with the Korea Food Codex's intestinal hemorrhagic E. coli microbiological test method

Products applied with the Food Standards Microbiological Test Method of Vero toxin producing E. coli (VTEC, Vero toxin producing E. coli)
High real-time PCR equipment versatility of master mix type
Application of UNG system to prevent false positive detection
Implementation of PCR reaction validation through IPC system​

LiliF® VTEC Real-time PCR Kit (ver2.0) is a product that implements the Korea Food Codex's intestinal hemorrhagic E. coli microorganism test method that can specifically detect Vero toxin gene stx1/stx2 (revised in April 2022, food Standard ▶ Article 8. General Test Method ▶ 4. Microbial Test Method ▶ 4.16 Real-time PCR Method of Enterohaemorrhagic E. coli). It is a versatile master mix type product that can detect up to 10 copy/uL and 1 copy/uL of each target gene and can use a variety of real-time PCR equipment. LiliF® VTEC Real-time PCR Kit (ver2.0) is a product that maximizes performance improvement and PCR efficiency by optimizing the food industry's stx1/2 specific primer/probe with its RealMOD™Probe M2 2X qPCR mix (with UDG). Uracil DNA Glycosylase (UDG) was added to verify the reliability and validity of the test results, and the UNG system was applied to prevent false-positive results from cross-over contamination by PCR amplicons. Internal positive control (IPC) DNA that does not affect target detection and IPC-specific primers/probes are added to the product to confirm IPC detection regardless of target amplification, so you can check the effectiveness of the PCR reaction.


 

Applications
•​  For research Use Only, Not for use in diagnostic procedures.
•​  The Kit is developed, designed, and sold for research purpose only.
•​  In-process monitoring for the presence of Vero toxin producing E.coli.

Kit Contents
No.Contents50 Tests/Kit
12X PCR Mix560 μl x 1 tube
2VTEC Detection Sol.280 μl x 1 tube
3Positive Control25 μl x 3 tubes
4DNase/RNase Free Water1 ml x 1 tube
Technical Information

Performance

 


 

● Result of detection of target nucleic acid VT1 : 10 copy/uL, VT2 : 1 copy/uL Confirmation of detection limit 

 

 

 

 

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TroubleShooting Guide

Q : Is the product reflecting the Korean Food Codex's intestinal hemorrhagic E. coli microbiological test method?

 

A : This product reflects the Food Standards Code (April 2022 revision) ▶ Article 8. General Test Method ▶ 4. Microbial Test Method ▶ 4.16 Enterohaemorrhagic Escherichia coli (2) Real-time PCR method. The target gene and specific primer/probe sequence information are the same as those listed, and the real-time PCR reaction conditions are also the same.​ 

 

 

 

Q : The detection signal is confirmed in the negative control test group.

 

A : This product contains primers and probes that specifically bind to Vero toxin (VT) generating genes (stx1, stx2). If the target amplification signal is confirmed in the negative control solution (DNase/RNase free water) provided in the product, retest in a situation where contamination is suspected.​ 

 

 

Q : Target detection is confirmed, but the IPC amplified signal is not.

 

A : If the concentration of nucleic acid in the sample used (genomic DNA extracted from the culture medium) is high, the reaction of a primer/probe specific to IPC DNA may affect IPC detection due to the competitive reaction of PCR. If target detection is confirmed, PCR is operating normally even if the IPC amplification signal is not confirmed, and it is okay to judge positive for the target gene. (However, if the IPC amplification signal is not confirmed even in the test result of the negative control solution provided in the product, please contact the manufacturer.)​ 

 

 

Q : Do I need to use specific real-time PCR equipment to use my product?

 

A : This product is a master mix type product consisting of 2x PCR mix and a primer/probe mixture (Detection solution). Therefore, it is recommended to completely thaw the product before use and simply centrifuge it before use. After mixing the required amount of PCR reaction solution according to the number of samples required, it is a product that can be used by dispensing it into a tube suitable for real-time PCR to be tested. This product can be used in real-time PCR equipment of​ 

 

 

Q : Is pretreatment necessary for test samples? Are there any DNA extraction products recommended by this product?

 

A : This product detects using DNA extracted from the enterohaemorrhagic E. coli enriched culture medium, and pre-treatment is essential. The extraction method verified through this product is the Boiling Method method presented in the Food Codex and our automatic nucleic acid extraction system (Miracle-AutoXT Automated Nucleic Acid Extraction system, IMC-NC15PLUS, extraction reagent: AutoXT total gDNA Extraction Kit, Cat No. 17188) -48/96), and the detection of the target gene was confirmed normally through two extraction methods. We recommend that you use our automated nucleic acid extraction system, which allows for simple and easy extraction of target nucleic acids.​ 

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