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time of measurements. After collection of all samples, serum copeptin levels were measured using “Human Copeptin ELISA Kits” (Phoneix Pharmaceuticals, USA; catalog no.: EK-065-32; lot no.: 603.858)” in microplate readers (Bio-Tek Instruments ELx800, USA
> G) and (Tru9I-rs757343 A > G), respectively Measurement of 25-(OH) D 3 The concentration of 25-(OH) D 3 was measured in the plasma of patients with MTC (n = 40) and those in the control group (n = 40) using an Elisa kit according to instructions by
Hepatitis E virus (HEV) strains are classified into 4 genotypes by nucleotide sequencing. Genotypes 3 and 4 infect humans and animals via HEV-contaminated food or water. HEV RNA was detected by PCR and antibodies were detected by ELISA. Since human studies showed that HEV IgG antibodies in sera can persist for extended periods, diagnosis of HEV infection in swine or humans is mainly based on serological detection using commercial ELISA kits. However, there is no supplemental method to verify ELISA results. Hence, we developed a novel method used for mutual correction of these common processes. Here, a modified stable HepG2 cell line was transfected with pcDNA3.1-ORF3 to express the swine HEV ORF3 protein. Based on this cell line, a novel immunoperoxidase monolayer assay (IPMA) was developed to detect antibodies against HEV. The results show that this method has good specificity, sensitivity and repeatability. When used to investigate 141 porcine serum samples, the IPMA had a coincidence rate of 92.2% with a commercial ELISA kit. The established IPMA described herein is valuable as a supplemental method to ELISA and can differentiate infections by HEV and other viruses.
Over the last few years several porcine epidemic diarrhoea (PED) outbreaks have been discovered in Europe including the first PED case in Slovenia in January 2015. The aim of this study was to determine when PED virus (PEDV) infection started in Slovenia. Serum samples collected between 2012 and 2016 were tested. Three hundred and seventy-five serum samples were collected from 132 Slovenian small, one-site pig farms. Samples were tested for PEDV antibodies utilising three different serological methods: commercially-available indirect ELISA, in-house blocking ELISA test and Immunoperoxidase Monolayer Assay (IPMA) test. One hundred and seventy (45.33%) tested samples were found positive by the commercially-available ELISA test kit, and 10 (5.68%) of these 170 samples found positive were positive by the in-house blocking ELISA. Only these 10 samples were collected from a farm where clinical signs of PED infection had been observed and PEDV was confirmed by RT-PCR methodology; the other 160 samples were collected randomly. Thirty-two samples with the highest S/P value obtained with the commercial ELISA were all negative with IPMA. Reasons for the high variance in the results obtained remain unclear; more research is required to ensure higher sensitivity and specificity in terms of PEDV antibody tests and other PED diagnostic methods.
FABP-9 was assessed by the Mybiosource ELISA test kit (cat no: MBS918033, sensitivity 0.047 ng/mL, reference range 0.188–12 ng/mL) obtained from MyBioSource, Inc., USA. The results were indicated as ng/mL and ng/mg protein. Total protein analysis was
enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer's instructions (Human Kisspeptin-10 Elisa Kit, standard curve range: 0.05 ng/mL-10 ng/mL, sensitivity: 0.02 ng/mL, intra-assay CV: 8%; interassay CV: 10%; catalog no. E3919Hu
ELISA kit of IBL (Hamburg, Germany). Basic level of cortisol was achieved by keeping off physical stress (physical activity, significant changes of environmental parameters as temperature, humidity, etc.) and emotional stress (fear, nervousness, panic
the middle (week #8), and at the end of the semester (week #16). Cortisol tests were performed using the procedure described by Sauvé et al. [ 26 ], by duplicate, following the SALIMETRICS-ELISA kits. Self-reported stress and eating behaviour
) and IL-6 were determined by enzyme-linked immunosorbent assay (ELISA). Briefly, blood was drawn from the peripheral vein of patients with MHD before dialysis or the healthy control on an empty stomach, in the morning. The blood was placed in a serum
Qualitative tests for C-reactive protein (CRP) are available for use in dogs, and provide a rapid in-house method of detecting acute inflammation. The aim of this study was to compare results from a qualitative CRP lateral flow test (Teco CRP FASTest) to those obtained from a quantitative CRP ELISA and to traditional methods of detecting inflammation, including total leukocyte and neutrophil numbers, presence of immature neutrophils and a left shift, presence or absence of toxic changes in neutrophils and plasma fibrinogen concentration in whole blood and serum samples collected from 113 client-owned dogs. More dogs had CRP FASTest positive results than had quantitatively increased CRP (ELISA) or increases in traditional methods used for measuring inflammation. Few dogs had increases in markers of inflammation but no elevated CRP. The qualitative CRP FASTest was found to be a sensitive test for detecting increased CRP concentration and was positive more frequently than were traditional markers of inflammation.
