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!