EURL STANDARD OPERATING PROCEDURES (SOP) FOR ASF DIAGNOSIS

This section provided detailed standard operating procedures (SOPs) of ASF diagnostic methods validated and currently available at the EURL-ASF. The interpretation of the results is reported in each procedure.

SAMPLING PROCESSING

Processing of samples for African swine fever (ASF) diagnosis. SOP/CISA/ASF/SAMPLES/1/ Method accredited UNE-EN ISO/IEC 17025:2017.

 

AFRICAN SWINE FEVER VIRUS (ASFV) GENOME DETECTION

Procedure for the extraction of the ASF virus (ASFV) DNA. SOP/CISA/ASF/DNA EXTRACTION/1/ Method accredited UNE-EN ISO/IEC 17025:2017.

Detection of the ASFV genome by the OIE-conventional PCR, Agüero et al., 2003. SOP/CISA/ASF/PCR/1 Method accredited UNE-EN ISO/IEC 17025:2017.

Detection of the ASFV genome by the OIE- real time PCR (1), King et al., 2003. SOP/CISA/ASF/PCR/2 Method accredited UNE-EN ISO/IEC 17025:2017.

Detection of the ASFV genome by the OIE- real time PCR (2), Fernández et al., 2013 (Taqman probe ASFV-VP72P1 or UPL commercial probe). SOP/CISA/ASF/PCR/3. Method accredited UNE-EN ISO/IEC 17025:2017.

ASF ANTIBODY DETECTION TECHNIQUES

Detection of ASFV antibodies by the OIE-indirect ELISA test. SOP/CISA/ELISA/1. Method accredited UNE-EN ISO/IEC 17025:2017

Detection of ASFV antibodies by the commercial INGENASA blocking ELISA test (INGEZIM PPA COMPAC K3.). SOP/CISA/ELISA/2. Method accredited UNE-EN ISO/IEC 17025:2017.

Detection of ASFV antibodies by the OIE-Immunoblotting (IB). SOP/CISA/IB/1. Method accredited UNE-EN ISO/IEC 17025:2017.

Detection of ASFV antibodies by the OIE-immunoperoxidase test (IPT). SOP/CISA/IPT/1. Method accredited UNE-EN ISO/IEC 17025:2017.

PRODUCTION OF THE ASFV ANTIBODY REAGENTS.

Production of the ASFV cytoplasmic soluble antigen (Ag) used in the OIE-indirect ELISA test an in the OIE-Immunoblotting (IB) test. SOP/CISA/ASF-Ag/1

Production of the OIE-Immunoblotting (IB) strips for ASF antibody detection. SOP/CISA/ASF/IB/STRIPS/1

Preparation of the ASFV-coated 96-well plates for the OIE-Indirect immunoperoxidase test (IPT). SOP/CISA/ASF/IPT-PLATES/1

GENOTYPING OF ASFV ISOLATES

Procedure for the genotyping of ASFV isolates by the analysis of partial regions of the ASFV genome. SOP/CISA/ASF/GENOTYPING/1

ASFV ISOLATION, TITRATION AND IDENTIFICATION BY THE HAEMADSORBING (HAD) TEST IN SWINE PRIMARY CELL CULTURES.

ASFV isolation and identification by the HAD in swine peripheral blood monocytes (PBMs). SOP/CISA/ASF/VI/1

ASFV isolation and identification by the HAD in porcine alveolar macrophages (PAMs). SOP/CISA/ASF/VI/2

ASFV GROWING AND TITRATION ON ESTABLISHED CELL LINES.

Procedure for growing and preparation of established cell lines for ASFV propagation (VERO, MS). SOP/CISA/ASF/CELLS/1

Procedure for ASFV growing and titration in established cell lines (VERO, MS). SOP/CISA/ASF/TITRATION/1