AccuPower® Salmonella spp. 3-Plex PCR Kit provides one-step detection of 2 serotypes (S. Choleraesuis and Typhimurium) and InvA common gene existed in Salmonella spp. 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® Salmonella
spp. 3-Plex PCR Kit |
Animal Disease Standard Diagnostic Method (QIA*) |
G Company (Germany) |
N Company (Canada) |
| InvA |
+ (Detected) |
+ (Detected) |
+ (Detected) |
+ (Detected) |
| S. Choleraesuis |
- (Not Detected) |
- (Not Detected) |
- (Not Detected) |
| S. Typhimurium |
| Notes |
Able to diagnose serotypes |
Unable to diagnose serotypes |
* 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 2 serotypes ST (S. Typhimurium) & SC (S. Choleraesuis) and Salmonella spp. common gene (InvA) through one 2-step PCR reaction in a tube.
> The optimized sensitivity and specificity
Confirmed on the 111 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 svstemic organs |
| Time to result |
1h 45min (Post-extraction) |
| Tests per Kit |
96 |
Performances
Figure 1. Electrophoresis of PCR product by AccuPower® Salmonella spp. 3-Plex PCR kit.
Using AccuPower® Salmonella spp. 3-Plex PCR Kit, Total of 4 ng template DNA [ATCC 7001 (S. Choleraesuis), ATCC 14028 (S. Typhimurium) 2 ng each)] was subjected to 95°C 5 min., 95°C 30 sec. 65°C 1 min 30 sec (30 cycles), 72°C 5 min PCR. 3 ul of amplified PCR product was ran on 2% TBE Agarose gel. Lane 1:
S.Typhimurium genomic DNA
Lane 2:
S. Choleraesuis genomic DNA
Lane 3:
S. Typhimurium &
Choleraesuis genomic DNA
Lane 4: S. Enteritidis genomic DNA
Lane M: 100 bp 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® Salmonella spp. 3-Plex PCR Kit and standard diagnostic method of QIA was compared by PCR using total of 4 ng template DNA [ATCC 7001 (S. Choleraesuis), ATCC 14028 (S. Typhimurium) 2 ng each)] under optimized condition: 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: AccuPower® Salmonellaspp. 3-Plex PCR Kit
Lane 2: Standard diagnostic method of QIA (Animal, Plant and Fisheries Quarantine and Inspection Agency, Korea)
Lane M: 100 bp DNA Ladder (D-1030)
Lane N: Negative control
Figure 3. Sensitivity test.
AccuPower® Salmonella spp. 3-Plex PCR Kit sensitivity test using total of 4 ng template DNA [ATCC 7001 (S. Choleraesuis), ATCC 14028 (S. Typhimurium) 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: 100 bp 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® Salmonella spp. 3-Plex PCR Kit, standard diagnostic method of QIA (Animal, Plant and Fisheries Quarantine and Inspection Agency, Korea), and commercially available diagnostic kit (I and K company). Total of 4 ng template DNA was subjected to PCR under optimized condition : 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: Wild -type S. enteritidis
Lane 2: Wild -type S. Rissen
Lane 3: Wild -type S. enteritidis
Lane 4: Wild -type S. Montevideo
Lane 5: Wild -type S. Schwarzengrund
Lane 6 ~ 12: Wild -type S. Typhimurium
Lane 13: Wild -type S. Cholerasuis
Lane 14: Wild -type S. Gallinarum
Lane 15: Wild -type S. Derby
Lane M: 100bp DNA Ladder (D-1030)
Lane N: Negative control
Figure 5. Specificity test against other entero-bacteria.
AccuPower® Salmonella spp. 3-Plex PCR Kit specificity against other entero-bacteria. Using AccuPower® Salmonella spp. 3-Plex PCR Kit, total of 4 ng template DNA was subjected to PCR 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.
(A)AccuPower® Salmonella spp. 3-Plex PCR Kit, (B)Standard diagnostic method of QIA (Animal, Plant and Fisheries Quarantine and Inspection Agency, Korea), (C)K company diagnostic kit.
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 : 100 bp DNA Ladder (D-1030)
Lane N : Negative control
