BackgroundSince 2010, outbreaks of genotype 2 (G2) porcine epidemic diarrhea virus (PEDV) have caused high mortality in neonatal piglets and have had devastating impacts on the swine industry in many countries. A reliable serological assay for evaluating the PEDV-specific humoral and mucosal immune response is important for disease survey, monitoring the efficacy of immunization, and designing strategies for the prevention and control of PED. Two PEDV spike (S) glycoprotein-based indirect enzyme-linked immunosorbent assays (ELISAs) were developed using G2b PEDV-Pintung 52 (PEDV-PT) trimeric full-length S and truncated S1–501 proteins derived from the human embryonic kidney (HEK)-293 cell expression system. The truncated S1–501 protein was selected from a superior expressed stable cell line. The sensitivity and specificity of these two ELISAs were compared to immunostaining of G2b PEDV-PT infected cells and to a commercial nucleocapsid (N)-based indirect ELISA kit using a panel of PEDV negative and hyperimmune sera.ResultsThe commercial N-based ELISA exhibited a sensitivity of 37%, a specificity of 100%, and a fair agreement (kappa = 0.37) with the immunostaining result. In comparison, the full-length S-based ELISA showed a sensitivity of 97.8%, a specificity of 94%, and an almost perfect agreement (kappa = 0.90) with the immunostaining result. Interestingly, the S1–501-based ELISA had even higher sensitivity of 98.9% and specificity of 99.1%, and an almost perfect agreement (kappa = 0.97) with the immunostaining result. A fair agreement (kappa< 0.4) was seen between the commercial N-based ELISA and either of our S-based ELISAs. However, the results of the full-length S-based ELISA shared an almost perfect agreement (kappa = 0.92) with that of S1–501-based ELISA.ConclusionsBoth full-length S-based and S1–501-based ELISAs exhibit high sensitivity and high specificity for detecting antibodies against PEDVs. Considering the high protein yield and cost-effectiveness, the S1–501-based ELISA could be used as a reliable, sensitive, specific, and economic serological test for PEDV.
Background The term big kidney‐little kidney syndrome in cats has been used for many years, but the definitions are not consistent and relevant research is limited. Objective To determine the factors that differ between normal and BKLK cats, as well as to develop models for predicting the 30‐day survival of cats with ureteral obstruction (UO). Animals Sixteen healthy cats and 64 cats with BKLK. Methods Retrospective study. To define BKLK by reference to data from clinically healthy cats. The demographic and clinicopathological data among groups were statistically analyzed. Results Big kidney‐little kidney syndrome cats had higher blood urea nitrogen (BUN) (median [interquartile range] 69 [28‐162] vs 21 [19–24] mg/dL, P < .001), creatinine (5.6 [1.9‐13.3] vs 1.3 [1.05‐1.40] mg/dL, P < .001), and white blood cells (10 800 [7700‐17 500] vs 6500 [4875‐9350] /μL, P < .001) and lower hematocrit (32.8 [27.1‐38.4] vs 39.1 [38.1‐40.4]%, P < .001), urine specific gravity (1.011 [1.009‐1.016] vs 1.049 [1.044‐1.057], P < .001) and pH (5.88 [5.49‐6.44] vs 6.68 [6.00‐7.18], P = .001) compared to the control cats. A lower body temperature (BT; 38.1 [37.9‐38.2] vs 38.7 [38.3‐39.2]°C, P = .009), higher BUN (189 [150‐252] vs 91 [36‐170] mg/dL, P = .04), and creatinine (15.4 [13.3‐17.4] vs 9.0 [3.1‐14.2] mg/dL, P = .03) were found among the UO cats that were not 30‐day survivors. A combination of BUN, phosphorus, and BT can predict 30‐day survival among UO cats with an area under receiver operating characteristic curve of 0.863. (P = .01). Conclusion An increase in the length difference between kidneys can indicate UO, but cannot predict outcome for BKLK cats.
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