Detection of Antibodies Against Canine Circovirus in Naturally and Experimentally Infected Canines by Recombinant Capsid Enzyme-Linked Immunosorbent Assay
Canine Circovirus (CanineCV), a new pathogen, found to be associated with hemorrhagic diarrhea dog, vasculitis, granulomatous lymphadenitis, and acute gastroenteritis. Although CanineCV is a very positive level in diarrhea cases, pathogenicity remains controversial. In this study, the prevalence and risk factors associated CanineCV infection among domestic dogs in northeastern China were investigated by enzyme-linked immunosorbent assay indirectly (iELISA) based on recombinant capsid protein.
The results revealed the proposed iELISA has no cross-reactivity with other related pathogens, and produce good diagnostic value. Then, to evaluate rCap iELISA, this study applied to the detection of antibodies against 1,047 CanineCV in clinical serum samples obtained from northeast China in 2016-2017. The results showed a positive level in five cities of Jilin, Liaoning and Heilongjiang provinces ranged from 22.22 to 42.29%. Statistical analysis showed a significant difference in age between the dog <3 months of age with respect to> dogs 1 year (p = 0.005), which is an infection that is more often identified CanineCV than an older dog.
In the experiment artificially infected dogs developed seroconversion after 9 or 12 days and the main way the virus is through faecal excretion. More interestingly, among the 32 ELISA positive serum samples, 34.75% of the samples tested positive for DNA CanineCV by qPCR, much higher than that in serum ELISA-negative samples (5.26%, 2/38). This report is the first to show that general CanineCV infection in the dog population in northeast China. The results showed a clear difference in the positive rate associated with diarrhea, age, but not with the different towns.
This study also provides a basis for evaluating the potential pathogens from CanineCV. But, pathogenicity, the relationship between antibody levels and immune protection, and the harmful effects of this virus remains to be established.
Detection of Antibodies Against Canine Circovirus in Naturally and Experimentally Infected Canines by Recombinant Capsid Enzyme-Linked Immunosorbent Assay
Standardized tests for enzyme-linked immunosorbent serosurvei of SARS-CoV-2 pandemic blood samples in clinical and home use
The extent of SARS-CoV infection-2 the entire population of the United States is currently unknown. High serology quality is a key tool for understanding the spread of infection, immunity to the virus, and correlates of protection. limited validation and testing of serological tests used to serosurvei can cause the data unreliable or misleading, and clinical trials using validated tests that can lead to misdiagnosis of expensive medical and public health decisions are not properly informed. Estimating the prevalence and clinical decision making depends on the specificity. Here, we present an optimized protocol based ELISA serology of antigen production data analysis.
This protocol defines the threshold for IgG and IgM seropositivity for the determination of the estimated specificity above 99%. Validation is done by using both traditional serum and dried blood collected in blood sample mail-in kit, use the archive (pre-2019) and known negative control PCR-positive patients diagnosed control. Minimal cross reactivity was observed for protein spikes of Mers, SARS1, OC43 and HKU1 virus and there is no cross-reactivity was observed with anti-influenza A H1N1 HAI titers during validation.
Description: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Rat CR. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat CR. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Rat CR, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Rat CR in the samples is then determined by comparing the OD of the samples to the standard curve.
Description: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Rat CR. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat CR. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Rat CR, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Rat CR in the samples is then determined by comparing the OD of the samples to the standard curve.
Description: A sandwich quantitative ELISA assay kit for detection of Human Calretinin (CR) in samples from serum, plasma, tissue homogenates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Human Calretinin (CR) in samples from serum, plasma, tissue homogenates or other biological fluids.
Description: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Human CR. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human CR. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Human CR, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Human CR in the samples is then determined by comparing the OD of the samples to the standard curve.
Description: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Human CR. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human CR. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Human CR, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Human CR in the samples is then determined by comparing the OD of the samples to the standard curve.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Calretinin (CR) in serum, plasma, tissue homogenates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Calretinin (CR) in serum, plasma, tissue homogenates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Calretinin (CR) in serum, plasma, tissue homogenates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Calretinin (CR) in serum, plasma, tissue homogenates and other biological fluids.
Description: Enzyme-linked immunosorbent assay based on the Double-antibody Sandwich method for detection of Human Calretinin (CR) in samples from serum, plasma, tissue homogenates and other biological fluids with no significant corss-reactivity with analogues from other species.
Description: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Rabbit CR protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rabbit CR. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Rabbit CR in the samples is then determined by comparing the OD of the samples to the standard curve.
Description: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Rabbit CR protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rabbit CR. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Rabbit CR in the samples is then determined by comparing the OD of the samples to the standard curve.
This strategy is very specific and is designed to give a good estimate of the seroprevalence of SARS-CoV-2 seropositive in a population, provide specific and reliable data from serosurvei and clinical trials that can be used to further evaluate and understand the SARS-CoV-2 immunity and correlates of protection.
