IDENTIFICAZIONE DI NUOVI BIOMARKER E METODI ANALITICI INNOVATIVI IN PATOLOGIA CLINICA VETERINARIA

Diagnostic test validation is challenge for the clinical pathologist when a new instrument or a new analyzer is introduced in the lab. The aim of this work is to assess the analytical and/or clinical performances of novel biomarker to evaluate a possible application as a useful marker in veterinary medicine. Four different objectives are investigate: 1. Haematological parameters: two new gates are draw, in particular a “high fluorescent region” to identify canine blood samples with cell of probable neoplastic origin that need further investigation and a gate to identify “high fluorescent platelet fraction” in macrothrombocytopenic Norfolk terrier. Both gates are useful to perform a correct diagnosis with a better outcome for the patients. 2. Inflammatory markers: the measurement of dROMs (derivative of reactive oxygen metabolites) in serum of dogs with leishmaniasis showe a strong involvement of these compounds in the symptomatic or the asymptomatic Leishmania infections, and could be use to identify dogs with sub-clinical forms. Another studies involve the validation of a new test for IFN-γ measurement after incubation of feline leukocytes with immunogenic peptides. This study is from one side a draw up of a new test with consequently assessment of analytical performances, and from another side a pathogenic study about FIP infection in cats. The paraoxonase 1 activity in dogs is the last inflammatory maker investigated, and results show a probable biological behaviour as a negative acute phase protein. 3. Hypertension markers: homocysteine and entothelin-1 (ET-1). Hypertension usually is secondary to diseases that indirectly could affect the systemic pressure, mainly renal and heart diseases. Homocysteine appears increased in dogs with both heart and kidney disease, but the clinical usefulness and the sensibility is not enough for positive impact on patient management. ET-1 appeared with higher sensibility and specificity in dogs with chronic kidney diseases and with hypertension, and its related with the severity of disease and could be a useful marker for the monitoring the follow up. 4. Development of new methods to investigate sialic acid characteristics and to assess if the different molecules bind to sialic acid molecules could interact with the development of the pathology. Results show an increased metastatic potential when sialic acid has α2,6 conformation in dogs with mammary tumour, and in cats the total sialic acid serum concentration is related to the immune response of cats during Coronavirus infections. 5. At last, evaluation of analytical performances of urine protein-to-creatinine ratio: it’s a commonly used test, but this study revealed a high imprecision that could affect test results.