AccuPower® ETEC-Pili 5-Plex PCR Kit provides one-step detection of 5 types of pilis produced by ETEC E. coli in animals: F4 (K88), F5 (K99), F6 (987P), F18 and F41. This featured kit is comprised of all the essential requirements for PCR reactions including primer, salts, dNTPs, and stabilizers in one tube to the provivded ready-to-use Premix format product. Users only need to add template DNA and D.W to perform the test.
| |
AccuPower® ETEC-Pili
5-Plex PCR Kit |
Animal Disease Standard
Diagnostic Method (QIA) |
Other Kits |
| F4(K88) |
+ (Detected) |
+ (Detected) |
Not Identifed same (similar) kit |
| F5(K99) |
| F6(987P) |
| F18 |
| F41 |
| Notes |
PreMix dried kit,
Detection limit of 104 cfu/ml |
Master solution Mix |
* QIA: Animal, Plant and Fisheries Quarantine and Inspection Agency, Korea
Features
Developed cooperatively with QIA (Quarantine and Inspection Agency), Application of patent completed (Patented).
> Fast response
Less than 2-hr detection of 5 pili-type pathogenic E. coli [F4 (K88), F5 (K99), F6 (987P), F18 & F41] 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.
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- Pili 5-Plex PCR kit.Using AccuPower® ETEC- Pili 5 Multiplex PCR Kit, Total of 5 ng template DNA (EcL1033/ EcL7805 / EcL2316 / EcL13316) 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-Pili genomic DNA
Lane N: Negative control
Lane M: 100 bp DNA Ladder (D-1030)
Figure 2. Comparison with standard diagnostic method of QIA (Animal, Plant and Fisheries Quarantine and Inspection Agency, Korea).AccuPower® ETEC- Pili 5-Plex PCR Kit and standard diagnostic method of QIA was compared by PCR using total of 5 ng template DNA (EcL1033/ EcL7805 / EcL2316 / EcL13316) under optimized condition: 95°C 5min., 95°C 30sec. 65°C 1min 30sec (30 cycles), 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-Pili 5-Plex PCR Kit
Lane 2: Standard diagnostic method of QIA
Lane N: Negative control
Figure 3. Sensitivity test.
AccuPower® ETEC-Pili 5-Plex PCR Kit sensitivity test using total of 5 ng template DNA (EcL1033/ EcL7805 / EcL2316 / EcL13316) 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: 100 bp DNA Ladder (D-1030)
Lane N: Negative control
Figure 4. 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-Pili 5-Plex PCR Kit, standard diagnostic method of QIA. Total of 5 ng template DNA was subjected to PCR under optimized condition featured product: 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. (A)AccuPower® ETEC-Pili 5-Plex PCR Kit PCR Kit, (B)Standard diagnostic method of QIA. Lane M: 100bp 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- Pili 5-Plex PCR Kit, total of 5 ng template DNA was subjected to PCR under: 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.
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: Acinetobacter 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: 100bp DNA Ladder (D-1030)
Lane N : Negative control
