Analytical quality assessment and method comparison of two immunoassays for the measurement of serum cardiac Troponin I in dogs and cats.

BACKGROUND: Cardiac troponin I (cTnI) is a sensitive and specific biomarker for myocardial injury. Validation of point-of-care (POC) analyzers for cTnI measurement is valuable to the critical care setting, in which rapid results can facilitate prompt diagnoses. An immunoassay for detecting cTnI is available for the POC AIA-360 analyzer (Tosoh Bioscience), but this has not been validated using canine and feline serum. OBJECTIVES: The objectives were (a) to determine precision, accuracy, and linearity of cTnI measurement using the AIA-360 immunoassay in pooled canine and feline samples, and (b) to compare results for individual canine and feline samples with those obtained using a reference chemiluminescence method (Immulite 1000, Siemens). METHODS: Intra- and inter-assay repeatability was determined using pooled canine and feline samples, and the coefficient of variation (CV) was calculated for each. Pooled samples were also serially diluted to assess linearity. A modified spike and recovery analysis was performed by mixing pooled samples with different concentrations. Bland-Altman and Deming regression analyses were used to determine bias for individual samples, and the total observed error (TEobs ) was calculated. RESULTS: Coefficient of variation values were well within the required maximum of 20%. Linearity was demonstrated over the range of samples tested, and the recovery study showed minimal proportional inaccuracies. Although the correlation between the analyzers was excellent, there was a large mean bias due to relative proportional bias. Total observed error consequently exceeded the total allowable error (TEA ). CONCLUSION: Although, in most respects, the analyzer demonstrated adequate performance, pronounced bias contributed to the large TEobs , indicating a requirement for analyzer-specific reference intervals.

Prevalence and risk factor analysis for feline haemoplasmas in cats from Northern Serbia, with molecular subtyping of feline immunodeficiency virus.

OBJECTIVES: The objectives of this study were to estimate the prevalence of feline haemoplasma infections in Northern Serbia, identify potential risk factors and perform molecular subtyping of feline immunodeficiency virus (FIV). METHODS: PCR analysis for feline haemoplasmas was performed on surplus EDTA blood samples from 373 cats from the Belgrade region, Serbia. An ELISA was used to determine the prevalence of feline leukaemia virus (FeLV) and FIV; PCR was performed on a subpopulation of these cats. FIV subtyping was performed using PCR. RESULTS: Within this population, 64/373 cats (17.2%) were infected with one or more haemoplasma species. Mycoplasma haemofelis was detected in 20/373 cats (5.4%), 'Candidatus Mycoplasma haemominutum' in 47/373 cats (12.6%) and 'Candidatus Mycoplasma turicensis' in 23/373 cats (6.2%). Coinfections were observed in 21/373 cats (5.6%). Based on ELISA serological retroviral testing, 4/310 cats (1.3%) were infected with FeLV, whereas 78/331 (23.6%) were infected with FIV. Multivariable analysis identified significant associations between haemoplasma infection and anaemia (anaemic/non-anaemic, odds ratio [OR] 2.7, 95% confidence interval [CI] 1.04-7.1; P = 0.041]), male gender (male/female, OR 4.5, 95% CI 2.22-9.03; P <0.0005), outdoor access (yes/no, OR 5.2, 95% CI 2.28-11.92; P <0.0005), non-pedigree breed (non-pedigree/pedigree, OR 5.5, 95% CI 1.24-24.84; P = 0.025) and FIV seropositive status (positive/negative, OR 2.4, 95% CI 1.21-4.83; P = 0.012). PCR analysis of the FIV ELISA-positive samples revealed clade D as being the most prevalent. CONCLUSIONS AND RELEVANCE: All three known species of feline haemoplasma were detected, confirming their presence in Serbia; 'Candidatus Mycoplasma haemominutum' was the most prevalent. We found a high prevalence of FIV-infected cats and FIV clade D was most prevalent.

Prevalence study and risk factor analysis of selected bacterial, protozoal and viral, including vector-borne, pathogens in cats from Cyprus.

BACKGROUND: Feline infectious agent studies are lacking in Cyprus. The aims of this study were to determine the prevalence and risk factors for various feline infectious agents, including feline vector-borne pathogens (FVBP), in cats from Cyprus. METHODS: A cross-sectional, descriptive, multicentre study was performed on 174 feline samples [138 owned and 36 shelter-feral, including both healthy (43) and non-healthy (131), cats] from private veterinary clinics from all six districts of Cyprus. Real-time quantitative polymerase chain reaction (qPCR) assays were used to detect Mycoplasma haemofelis (Mhf), "Candidatus Mycoplasma haemominutum" (CMhm) and "Candidatus Mycoplasma turicensis" (CMt). The population was tested for four FVBP including Bartonella henselae and Leishmania spp. using qPCR, while conventional PCR assays were used to detect Ehrlichia/Anaplasma spp. and Hepatozoon spp. Serological assays were performed to detect Leishmania infantum antibodies, feline leukaemia virus (FeLV) antigen and feline immunodeficiency virus (FIV) antibodies. Statistical analysis was performed to test associations and possible risk factors between variables and infectious agents. RESULTS: Ninety-six (55.2%) of the 174 cats were PCR-positive for at least one infectious agent. Forty-six cats (26.4%) were haemoplasma positive, including 13 (7.5%) for Mhf, 36 (20.7%) for CMhm and 12 (6.9%) for CMt. Sixty-six cats (37.9%) were positive for Hepatozoon spp., while 19 (10.9%) were positive for B. henselae, four (2.3%) for Leishmania spp. and one (0.6%) for Ehrlichia/Anaplasma spp. Sequencing revealed the presence of Hepatozoon felis, L. infantum and Anaplasma platys. Of the 164 cats that underwent retroviral serology, 10 (6.1%) were FeLV-positive and 31 (18.9%) were FIV-positive, while L. infantum serology was positive in 7 (4.4%) of the 160 cats tested. Multivariable logistic regression revealed significant associations for various infectious agents including L. infantum with each of Hepatozoon spp. and CMt infection. CONCLUSIONS: A high prevalence of infectious agents was found in cats from Cyprus with Mhf, CMhm, CMt, L. infantum, B. henselae, H. felis, A. platys, FeLV and FIV infections reported for the first time. The significant associations between different pathogens provide a better understanding of similarities in the epidemiology of these pathogens and interactions between them.