Upload
sergio-exopol
View
158
Download
2
Tags:
Embed Size (px)
Citation preview
Real Time PCR assays for veterinary diagnosis
Real Time PCR (qPCR) assays Use of qPCR assays for diagnosis of animal diseases have increased
considerably: Better sensitivity and specificity. Short time results. Capable of quantification.
Drawbacks of most commercial qPCR assays:
Several reagents: more variability inter-assay and possibility of mistakes.
Different designs: each qPCR has a different setup / thermal profile. Poor validation: “Research use only”
Liquid format“OneMIX” Real Time PCR assays
Stabilized formatReal Time PCR assays
All qPCR reagents in a single tube. A novel technology allows stabilization of all reagents into a qPCR plate.
Pathogen and EC PRIMERS
Endogenous control’s
PROBE (HEX)
Pathogen’sPROBE (FAM)
Enzyme
BUFFERS & dNTPs
Shipment/Store at 4°C.
Shipment/Store at -20°C.
Make qPCR easy
1. Select the required number of reactions.
2. Add 15µl nucleic acid free water/each well.
4. Run your qPCR.
Get results in less than 2 hours!
3. Add 5µl of DNA sample / PC/ NC.
2. Just add 15µl of oneMIX reaction in each tube/well.
Other advantages:
Avoid false negative results: endogenous control (EC) in each well.
Pathogen detection (FAM)
EC detection (HEX)
Relative or absolute quatification: specific synthetic positive controls.
Same thermal protocol: test several pathogens in a single run.
Pathogen APathogen B
Pathogen C
Multiparametric qPCR assays for Porcine abortion panel
PCV2
PPV
C. abortus
C. suis
Leptospira
SAMPLE
Multiparametric qPCR assays for E. coli virulence factors
Edema disease(ED)
Post-weaning diarrhea(PWD)
Stx2e F18 F4 Sta Stb LT EC
SAMPLE
Conveniently validated:
• Reference strain, vaccine strain, field strains.
• 42 other related pathogens.
Sensitivity and Specificity
• Reportanble range: using specific positive controls.
• Intra-assay e inter-assay studies.
Limits of detection (LOD)
Precision
• Concordance study: commercial qPCR assay or other diagnostics test.Accuracy
• Different species.• Several type of samples.Clinical cases
Stabilized qPCR assays
PATHOGENS
1Bovine Herpesvirus
12 Mycoplasma bovis
3Pathogenic Leptospira
4 Toxoplasma gondii
5 Coxiella burnetii
6 Neospora caninum
7Tritrichomonas
foetus
Available “oneMIX” qPCR assays
BESN EXOone Besnoitia besnoitiHPS
UEXOone Haemophilus parasuis
MHYR
EXOone Mycoplasma hyorhinisEXOone Salmonella
entericaSENT
Available “oneMIX” qPCR assays
CRYP
EXOone Cryptosporidium spp
In testing phase:
BVDV/BDV
Bovine Respiratory Syncytial virus
Bovine Parainfluenza 3
RHDV new variant
Thank you
for your
attention!