AccuPower® ETEC-Toxin 4-Plex PCR Kit

AccuPower® ETEC-Toxin 4-Plex PCR Kit is designed to detect 4 types of E. coli toxin gene simultaneously such as LT, STa, STb and STX2e

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Cat. No.

MAT-1115

Product Overview

AccuPower® ETEC-Toxin 4-Plex PCR Kit provides one-step detection of 4 types of toxins produced by ETEC E. coli in animals: LT, STa, STb, and STX2e. This featured kit is comprised of all the essential requirements for PCR reactions including primer, salts, dNTPs, and stabilizers in one tube to provide the ready-to-use premix format product. Users only need to add template DNA and D.W to perform the test.

	AccuPower® ETEC-Toxin 4-Plex PCR Kit	Animal Disease Standard Diagnostic Method (QIA)	G Company (Germany)
LT	+ (Detected)		+ (Detected)
STa			- (Not detected)
STb			- (Not detected)
STX2e			- (Not detected)
Notes	PreMix dried kit, Detection limit of 10⁴ cfu/ml	Master solution Mix	Master solution Mix

* QIA: Animal, Plant and Fisheries Quarantine and Inspection Agency, Korea

Features

Developed cooperatively with QIA, and Application of patent completed (Patented)

> Fast response
Less than 2-hr detection of 5 types of toxins (LT, STa, STb and STX2e) produced by ETEC E. coli in animals through one 2-step PCR reaction in a tube.

> The optimized sensitivity and specificity
Confirmed on the 171 in-field strains owned by QIA. Application of patented Enzyme-mediated HotStart technology: 1x10⁴ CFU/ml of optimized sensitivity, possible exclusion of false potisive.

> Great reproducibility
Dried-type HotStart PCR PreMix type – Addition of template DNA and DW only required & highly reproducible result provided.

> Prevention of amplicon-induced cross-contamination by 8-MOP
Binding of 8-MOP (8-methoxypsoralen) to DNA double-helix structure, followed by UV irradiation.

> High Stability
1-year warranty under -20 °C.

> Quality Control

All AccuPower® products are manufactured under ISO 9001 Quality Control System.

Technical/Specs

Specifications

Instrument	AllInOneCycler™ Thermal Block or other Thermal Block
Specimen	Feces, small and large intestines, septicemia, urine, other systemic organs
Time to result	1h 45min (Post-extraction)
Tests per Kit	96

Performances

Figure 1. Electrophoresis of PCR product by AccuPower® ETEC-Toxin 4-Plex PCR kit
Using AccuPower® ETEC-Toxin 4-Plex PCR Kit, Total of 4 ng template DNA (EcL6601/EcL7805, 2ng each) was subjected to 95°C 5min., 95°C 30sec. 65°C 1min 30sec (30cycles), 72°C 5min PCR. 3 ul of amplified PCR product was ran on 2% TBE Agarose gel.
Lane 1: ETEC-Toxin genomic DNA
Lane M: 100bp DNA Ladder (D-1030)
Lane N: Negative control

Figure 2. Comparison with standard diagnostic method of QIA (Animal, Plant and Fisheries Quarantine and Inspection Agency, Korea).
AccuPower® ETEC-Toxin 4-Plex PCR Kit and standard diagnostic method of QIA was compared by PCR using total of 4 ng template DNA (EcL6601 / EcL7805, 2ng each) under optimized condition: 95°C 5min., 95℃ 30sec. 65°C 1min 30sec (30cycles), 72°C 5min. 3 ul of amplified PCR product was ran on 2% TBE Agarose gel.
Lane M: 100 bp DNA Ladder (D-1030)

Lane 1: AccuPower® ETEC-Toxin 4-Plex PCR Kit

Lane 2: Standard diagnostic method of QIA

Lane N: Negative control

Figure 3. Sensitivity test.
AccuPower® ETEC-Toxin 4-Plex PCR Kit sensitivity test using total of 4 ng template DNA (EcL6601^TM/ EcL7805^TM 2 ng each) under: 95°C 5 min., 95°C 30 sec. 65°C 1 min 30 sec (30 cycles), 72°C 5 min. 3 ul of amplified PCR product was ran on 2% TBE Agarose gel.
Lane 1: 1X10⁸ CFU/ml
Lane 2: 1X10⁷ CFU/ml
Lane 3: 1X10⁶ CFU/ml
Lane 4: 1X10⁵ CFU/ml
Lane 5: 1X10⁴ CFU/ml
Lane 6: 1X10³ CFU/ml
Lane 7: 1X10² CFU/ml
Lane M: 100bp DNA Ladder(D-1030)
Lane N: Negative control

Figure 4. Specificity comparison with standard diagnostic of QIA (Animal, Plant and Fisheries Quarantine and Inspection Agency, Korea) and commercially available diagnostic kit.
24 field strains were tested using AccuPower® ETEC-Toxin 4-Plex PCR Kit, standard diagnostic method of QIA and commercially available diagnostic kit (K company). Total of 4 ng template DNA (EcL6601 / EcL7805, 2ng each) was subjected to PCR under optimized condition: 95°C 5min., 95°C 30sec. 65°C 1min 30sec (30cycles), 72°C 5min. 3 ul of amplified PCR product was ran on 2% TBE Agarose gel.
(A)AccuPower® ETEC-Toxin 4-Plex PCR Kit PCR Kit, (B)Standard diagnostic method of QIA, (C)K company diagnostic kit

Lane M: 100 bp DNA Ladder (D-1030)

Lane 1~24: Wild -type DNA (field strains)

Lane N: Negative control

Figure 5. Specificity test against other entero-bacteria.
Using AccuPower® ETEC-Toxin 4-Plex PCR Kit, total of 4 ng template DNA was subjected to PCR under: 95°C 5min., 95°C 30sec. 65℃ 1min 30sec (30cycles), 72°C 5min. 3 ul of amplified PCR product was ran on 2% TBE Agarose gel.
Lane 1: Acinetobacter lwoffii genomic DNA

Lane 2: Enterococcus fecalis genomic DNA

Lane 3: Maltophilia genomic DNA

Lane 4: Pseudomonas putida genomic DNA
Lane 5: Proteus mirabilis genomic DNA

Lane 6: Citrobacter freundii genomic DNA

Lane 7: Klebsiella pneumonia sub. genomic DNA

Lane 8: Enterobacter cloacae genomic DNA

Lane 9: Acinetobac-ter haemolyticus genomic DNA

Lane 10: Comamonas testosterone genomic DNA

Lane 11: Clostridium perfrigens alpha genomic DNA

Lane 12: Burkholderia cepacia genomic DNA

Lane 13: Ochrobactrum anthropi genomic DNA

Lane 14: Serratia marcescens genomic DNA
Lane 15: Acinetobacter junii genomic DNA

Lane M: 100 bp DNA Ladder (D-1030)

Lane N : Negative control

Ordering Info

Ordering information

Cat. No.	Product Description	Price
MAT-1115	AccuPower^® ETEC-Toxin 4-Plex PCR Kit	Inquire

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Quality assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

ISO 9001 - certificate

ISO 13485 - certificate