Jane H C Huang, Bianca N Lourenço, Canaan Whitfield-Cargile
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引用次数: 0
Abstract
Objective: To develop and analytically validate a sandwich ELISA that measures angiotensinogen concentration in canine urine and serum samples.
Methods: Serum (n = 4) and urine (10) samples submitted to a clinical pathology laboratory were used for assay development. Recombinant canine angiotensinogen and both rabbit-derived polyclonal and mouse-derived monoclonal antibodies against it were obtained from a commercial source. A sandwich ELISA was developed and validated via assessment of precision, limit of blank, dilutional linearity, recovery, and cross-reactivity with albumin and angiotensin II.
Results: Intra- and interassay variability ranged from 8.31% to 14.52% and 1.88% to 16.87%, respectively. The limit of blank of the assay was 1.01 ng/mL, which was lower than all observed concentrations in 4 serum (27,130 to 64,690 ng/mL) and 10 urine (3.24 to 8,970.24 ng/mL) samples. Both serum and urine demonstrated ideal dilutional linearity following 2-fold serial dilutions (R2 > 0.99). The assay showed moderate recovery (61.8% to 113.9%). Minimal cross-reactivity was observed with albumin and angiotensin II, as both yielded values below the assay's limit of blank.
Conclusions: The angiotensinogen ELISA we developed is precise, linear, and sufficiently sensitive for measuring angiotensinogen in the urine and serum of domestic dogs.
Clinical relevance: Urinary angiotensinogen quantification is a promising noninvasive marker of intrarenal renin-angiotensin system activity in humans and rodents; this assay enables evaluation of its utility in dogs.
期刊介绍：
The American Journal of Veterinary Research supports the collaborative exchange of information between researchers and clinicians by publishing novel research findings that bridge the gulf between basic research and clinical practice or that help to translate laboratory research and preclinical studies to the development of clinical trials and clinical practice. The journal welcomes submission of high-quality original studies and review articles in a wide range of scientific fields, including anatomy, anesthesiology, animal welfare, behavior, epidemiology, genetics, heredity, infectious disease, molecular biology, oncology, pharmacology, pathogenic mechanisms, physiology, surgery, theriogenology, toxicology, and vaccinology. Species of interest include production animals, companion animals, equids, exotic animals, birds, reptiles, and wild and marine animals. Reports of laboratory animal studies and studies involving the use of animals as experimental models of human diseases are considered only when the study results are of demonstrable benefit to the species used in the research or to another species of veterinary interest. Other fields of interest or animals species are not necessarily excluded from consideration, but such reports must focus on novel research findings. Submitted papers must make an original and substantial contribution to the veterinary medicine knowledge base; preliminary studies are not appropriate.
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