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 (EcL6601TM/ EcL7805TM 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: 1X10
8 CFU/ml
Lane 2: 1X10
7 CFU/ml
Lane 3: 1X10
6 CFU/ml
Lane 4: 1X10
5 CFU/ml
Lane 5: 1X10
4 CFU/ml
Lane 6: 1X10
3 CFU/ml
Lane 7: 1X10
2 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 kitLane 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
