OIE Reference Laboratory for Babesiosis and Theileriosis. National Reference Centre for Anaplasma, Babesia, Rickettsia and Theileria (C.R.A.Ba.R.T.)

Pathogen DNA detection is carried out on DNA extracted from blood, tissue, cell cultures and arthropod samples. DNA extraction can be performed starting from fresh or frozen (– 20°C ) blood and tissues (spleen is the most suitable organ for pathogen detection). Arthropods (ticks or fleas) fresh or stored in 70% alcohol or in nucleid acid preserving reagent “RNA later “ at room temperature can be analysed.

PCRs are performed to detect DNA of pathogens belonging to Anaplasma, Babesia, Rickettsia  and Theileria genera. Real Time PCRs are carried out for A. marginale, B. bovis, B. bigemina and Rickettsias spp. Reverse line blotting is carried out for Babesia and Theileria species in ruminants. Sequencing is carried out to confirm diagnosis and for species/strain typing.
Details of the exams are reported in the tables below.

The Standard Operative Procedures (POS/PdP) are certified according to UNI CEI EN ISO/IEC 17025:2005.

Molecular diagnostic

Pathogen Matrix Method POS/PdP
A. marginale Blood, tissue and arthropod samples Real-time PCR
Anaplasma spp Blood, tissue and arthropod samples PCR POS-CRA 22
A. marginale Blood, tissue and arthropod samples PCR POS-CRA 03
A. ovis Blood, tissue, and arthropod samples PCR
A .phagocytopylum Blood, tissue and arthropod samples PCR POS-CRA 11
All the pathogenes Purified PCR products Sequencing

 

Pathogen Matrix Method POS/PdP
B. bovis, B. bigemina Blood, tissue and arthropod samples Real-time PCR
B. bigemina Blood, tissue and arthropod samples PCR POS-CRA 02
B. bovis Blood, tissue and arthropod samples PCR POS-CRA 09
B. ovis, B. canis Blood, tissue and arthropod samples PCR
B. caballi Blood, tissue, and arthropod samples PCR POS-CRA 15
B. microti Blood, tissue, and arthropod samples PCR POS-CRA 21
B. bigemina, B. bovis, B. divergens, B. major, B. motasi, B. ovis, B. crassa Blood, tissue, and arthropod samples Reverse line blotting  

All the pathogenes Purified PCR products Sequencing

 

Pathogen Matrix Method POS/PdP
Rickettsia spp., Rickettsia conorii Blood, tissue and arthropod samples Real-time PCR
Rickettsia spp Blood, tissue and arthropod samples PCR POS-CRA 04
All the pathogenes Purified PCR products Sequencing

 

Pathogen Matrix Method POS/PdP
T.annulata Blood, tissue and arthropod samples PCR POS-CRA 01
T.ovis Blood, tissue and arthropod samples PCR
T.equi Blood, tissue and arthropod samples PCR POS-CRA 14
T. annulata, T. velifera, T. taurotragi,

 T. mutans,

T. buffeli/orientalis,

T. ovis, T. lestoquardi, T. hirci

 

Blood, tissue, and arthropod samples

Reverse line blotting  

 

All the pathogenes Purified PCR products Sequencing

Serological diagnostic

Pathogen Matrix Method POS/PdP
A. marginale Ruminants serum ELISA assay POS-CRA 07
B. bigemina Serum of animals receptive ELISA assay POS-CRA 18
A. phagocitophilum Serum of animals receptive Immunofluorescence assay POS-CRA 12
B. canis Dog serum Immunofluorescence assay POS-CRA 16
B. bovis Serum of animals receptive Immunofluorescence assay POS-CRA 20
B. microti Serum of animals receptive Immunofluorescence assay
B. caballi Horse serum Immunofluorescence assay POS-CRA 10
R. conorii Serum of animals receptive Immunofluorescence assay POS-CRA 05
T. equi Horse serum Immunofluorescence assay POS-CRA 13

Plasmid DNA testing

  Method
PCR products starting from any pathogen Cloning

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Browse Catalog Contact: Dr. Santo Caracappa santo.caracappa@izssicilia.it

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