ASF DIAGNOSIS TOOLS. Prof. José M. Sánchez- Vizcaíno Berlin, 2011 LABORATORY ASF REFERENCE

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1 ASF DIAGNOSIS TOOLS Berlin, 2011 ASF REFERENCE LABORATORY Prof. José M. Sánchez- Vizcaíno

2 ASFV: A old friend MAIN WORK: DIAGNOSIS and ERADICATION MODELS ASF REFERENCE LABORATORY

3 ASF

4 Agenda: A rapid View 1) LAB. DIAGNOSIS: a) Key points of Asf diagnosis b) Tests Available 2) ASF PROTECTION: Vaccine 3) EARLY DETECTION AND CONTINGENCY PLAN 4) THINGS TO DO

5 ASF Laboratory DIAGNOSIS: KEY POINTS No Vaccine Available Abs good infectious marker!!!! No Neutralizing Antibodies Antibodies = INFECTION Viremia for Long period of Time Antibodies Persist During Month, Even Years From dpi. SEVERAL TEST AVAILABLE. In Good Condition

6 ASF DIAGNOSIS. Key points INFECTION DISEASES/DEAD CARRIERS 12 h to 1 week 1 3 weeks 4 Weeks to 2 years VIRUS Ab Infection Clinical Carriers

7 ASF LABORATORY TESTs ASF LABORATORY TESTs Identification of the Agent Virus Isolation in cell cultures: TIME: 3 to 10 Days Haemoadsorption autorosette autorosette (HA) test with peripheral blood leukocytes from infected pigs. Mandatory in first notification in free country ONLY Reference Labs!!

8 ASF LABORATORY DIAGNOSIS ASF LABORATORY DIAGNOSIS DNA Detection TIME: 5 to 6 H PCR: CONVENTIONAL and REAL TIME 0 dpi 1 dpi 2 dpi 3 dpi 4 dpi 7 dpi B S B S B S B S B S B S M 257 bp- 257 bp- 257 bp- 108 bp- King et al., C M V Agüero et al., 2003 MOST COMMONLY USED PPC-3/4 + PPA- 1/2 Agüeroet al., 2004

9 ASF LABORATORY DIAGNOSIS ASF LABORATORY DIAGNOSIS Antigen Detection TIME 75 MINUTES Direct immunofluorescence (DIF) Easy to use Personal trained needed to interpretate the results NEGATIVE POSITIVE Low sensitivity in subacute and chronic forms Significant lack of sensitivity due to Ag-Ab Ab complex formation. Not recomended for analysis of serum and tissue homogenated samples after first week pi. due to false negative results. r

10 Antigen Detection Immunochromatography TIME 30 MINUTES Pen side tests Easy to use ASF LABORATORY DIAGNOSIS ASF LABORATORY DIAGNOSIS No special equipment needed PPA-CROM ANTIGEN DETECTION POSITIVE SAMPLE NEGATIVE SAMPLE

11 ASF LABORATORY DIAGNOSIS ASF LABORATORY DIAGNOSIS Antigen Detection Immunochromatography Good working with high levels of virus Lower sensibility than PCR ORIGINAL EASY TO USE RAPID RESULTS No training need C- C+ 10-2

12 ASF LABORATORY DIAGNOSIS ASF LABORATORY DIAGNOSIS Antigen Detection ELISA DAS TIME 3 HOURS Antigen detection in spleen samples INGEZIM PPA DAS

13 ASF LABORATORY Ab detection ASF LABORATORY Ab detection Antibody Detection ELISA tests Indirect ELISA (OIE) In House ELISAs Commercial ELISA, Ingezim PPA COMPACT MOST COMMONLY USED. TIME 3 H Indirect immunofluorescent test (IIF) Immunoblotting TIME 3 H CONFIRMATORY TEST

14 ASF DIAGNOSIS. Key points INFECTION DISEASES/DEAD CARRIERS 1 week 2 3 weeks 4 Weeks to 2 years C M V 257 bp C M V 257 bp- 108 bp- 108 bp- Infection Clinical Carriers

16 ASF PROTECTION: NO VACCINE NO INACTIVATED VACCINE NO ATENUATED VACCINE. ONLY PARCIAL PROTECTION NO RECOMBINAT VACCINE: NO TARGET GENES NO DNA Gene Delection Subunit: partial ANTIBODIES ARE RELATED WITH SOME TYPE OF PROTECTION AS WELL AS IN CHRONIC AND ENDEMIC ASF INFECTION Eradication without vaccine is possible but not easy. Spain

17 ASF. Spain: IN DOOR

18 ASF CONTROL EARLY DETECTION CONTINGENCY PLANS

19 ASF EARLY DETECTION:

20 FIELD TRAINING: Farmers and Vets

21 FARMERS INFORMATION: Risk points

22 MANUAL for a FAST RESPONSE 1. A good contingency plan adapted to the risk and scenario 2. NOTIFICATION SYSTEM 3. ZONING OF AFECTED AREAS 4. BAN ON ANIMALS MOVEMENTS 5. LABORATORY CONFIRMATION 6. PROCEDURE FOR DESTRUCTIONS OF CARCASSES 7. DEPOPULATION 8. CLEANING AND DISINFECTION 9. SEROLOGICAL CONTROLS 10.STUDY WILD BOAR AND/OR VECTORS 11.SENTINEL ANIMALS 12. REPOPULATION

23 Eradication s Key action: Our experience a)aproved programe with farmers. Good information b)early detection and good contingen plans c)detection of positives and carriers animals by serology d)elimination of all positives and carriers animals e)improvements of biosecurity in farms (in side and outside) f)include good Restrictions areas g)control of movements. Identifications h)control of ticks (Elisa for ticks) i)economical compensation

24 THINGS TO DO A good early detection program A good contingency plan, adapted to the Risk. A good control program adapted to the different scenarios. Better knowledge of ticks and WB: Control & distribution. Good collaboration. OIE Lab and our expertise is yours.

25 AGRADECIMIENTOS Carmen Bulboa Deborah Kukielka Facundo Linares Luis Martín Otero Beatriz Martínez Marta Martínez Lina Mur Ana C. Pérez Belén Rivera Rocío Sánchez Almudena Sánchez Víctor Rodríguez Raquel Vargas Marina Vicente

Diagnostic Testing and Strategies for BVDV

Diagnostic Testing and Strategies for BVDV Dan Grooms Dept. of Large Animal Clinical Sciences College of Veterinary Medicine Introduction Clinical diseases in cattle resulting from infection with bovine