14 research outputs found
Four Amino Acid Changes in HIV-2 Protease Confer Class-Wide Sensitivity to Protease Inhibitors
Vergleich der mechanischen Beanspruchung von Knietotalendoprothesen bei unterschiedlichen Knieverschleiß-Normen mit einem Finiten Elemente Modell
Validation of a human immunoturbidimetric assay for measuring canine C-reactive protein
C-reactive protein (CRP) is an extremely powerful prognostic
marker for a wide variety of diseases. In human medicine
immunoturbidimetric assays (ITAs) are replacing the ELISA test
due to their high degree of accuracy, high-throughput and low
turnaround time. Until now, a canine-specific ELISA test has
represented the gold standard in veterinary medicine for measuring
serum CRP. Even though there are contrasting data concerning the
cross-reactivity of human and canine CRP, a previous study has
nevertheless demonstrated that a commercially available human
ITA is reliable for measuring canine CRP without method
modification. Since many automated analyzers are furnished with
proprietary standardized reagents, the aim of this study was to
verify whether a commercially available human ITA (Olympus
system reagent, CRP ITA, Olympus, Italy) different from the
previous validated assay is also reliable for accurately measuring
canine CRP. The CRP concentrations of 35 canine sera collected from
either healthy dogs or dogs with infectious and neoplastic diseases
were assayed by ELISA and ITA on an automated chemistry
analyzer (Olympus AU 400). The ELISA test was carried out
following the manufacturer\u2019s instructions while the ITA measurement
was initially performed using the settings and calibration of
the human assay. The results, which were compared using Passing
and Bablok regression analysis, showed that the ITA underestimated
canine CRP both with an absolute and a proportional
inaccuracy. To overcome this pitfall, the ITA was repeated after
calibration of the automated analyzer with a standard specimen
obtained by pooling 3 canine sera with high CRP concentrations.
The standard was assayed 6 times in the same run using the ELISA
test. The CV was 5.8% and the mean concentration of the 6 replicates
was assumed to be the actual concentration of the standard. The
modified assay (ITA-c) was used to evaluate the same 35 serum
samples in duplicate. The results were again compared with the
ELISA test by Passing and Bablok regression analysis which showed
a good correlation (y51.18x \u2013 0.69). Moreover, two samples with
23.7 mg/dl and 21.7 mg/dl of CRP were serially diluted and
assayed in duplicate. The ITA-c was linear in the verified range
(y50.9988x \ufe 0.2 and y50.9982x \ufe 0.07, respectively). In conclusion,
this study confirms that any commercially available human
CRP ITA should be validated before its routine application in dogs.
Moreover, the partial cross-reactivity between human and canine
CRP allowed us to reliably measure canine CRP after calibration of
the ITA with a standard pooled sample having a known concentration
of canine CRP
Validation of a human immunoturbidimetric assay for measuring canine C-reactive protein
C-reactive protein (CRP) is an extremely powerful prognostic marker for a wide variety of diseases. In human medicine immunoturbidimetric assays (ITAs) are replacing the ELISA test due to their high degree of accuracy, high-throughput and low turnaround time. Until now, a canine-specific ELISA test has represented the gold standard in veterinary medicine for measuring serum CRP. Even though there are contrasting data concerning the cross-reactivity of human and canine CRP, a previous study has nevertheless demonstrated that a commercially available human ITA is reliable for measuring canine CRP without method modification. Since many automated analyzers are furnished with proprietary standardized reagents, the aim of this study was to verify whether a commercially available human ITA (Olympus system reagent, CRP ITA, Olympus, Italia) different from the previous validated assay is also reliable for accurately measuring canine CRP.
The CRP concentration of 35 canine sera collected from either healthy dogs or dogs with infectious and neoplastic diseases was assayed by ELISA and ITA on an automated chemistry analyzer (Olympus AU 400). The ELISA test was carried out following the manufacturer\u2019s instructions while the ITA measurement was initially performed using the settings and calibration of the human assay. The results, which were compared using Passing and Bablok regression analysis, showed that the ITA underestimated canine CRP both with an absolute and a proportional inaccuracy. To overcome this pitfall, the ITA was repeated after calibration of the automated analyzer with a standard specimen obtained by pooling three canine sera with high CRP concentrations. The standard was assayed 6 times in the same run using the ELISA test. The CV was 5.8% and the mean concentration of the 6 replicates was assumed to be the actual concentration of the standard. The modified assay (ITA-c) was used to evaluate the same 35 samples in duplicate. The results were again compared with the ELISA test by Passing and Bablok regression analysis which showed a good correlation (y = 1.18x \u2013 0.69). Moreover, two samples with 23.7mg/dl and 21.7 mg/dl of CRP, respectively, were serially diluted and assayed in duplicate. The ITA-c was linear in the verified range (y = 0.9988x + 0.2 and y = 0.9982x + 0.07).
In conclusion, this study confirms the evidence that any commercially available human CRP ITA should be validated before its routine application in dogs. Moreover, the partial cross-reactivity between human and canine CRP allowed us to reliably measure canine CRP after calibration of the ITA with a standard specimen having a known concentration of the analyte