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