scholarly journals Evaluation of ELISA tests for the qualitative determination of IgG, IgM and IgA to SARS-CoV-2

Author(s):  
Colavita Francesca ◽  
Brogi Alessandra ◽  
Lapa Daniele ◽  
Bordi Licia ◽  
Matusali Giulia ◽  
...  

AbstractSerological assays for anti-SARS-CoV-2 antibodies are now of critical importance to support diagnosis, guide epidemiological intervention, and understand immune response to natural infection and vaccine administration. We developed and validated new anti-SARS-CoV-2 IgG, IgM and IgA ELISA tests (ENZY-WELL SARS-CoV-2 ELISA, DIESSE Diagnostica Senese S.p.a.) based on whole-virus antigens. We used a total of 553 serum samples including samples from COVID-19 suspected and confirmed cases, healthy donors, and patients positive for other infections or autoimmune conditions. Overall, the assays showed good concordance with the indirect immunofluorescence reference test in terms of sensitivity and specificity. Especially for IgG and IgA, we observed high sensitivity (92.5 and 93.6%, respectively); specificity was high (>96%) for all antibody types ELISAs. In addition, sensitivity was linked to the days from symptoms onset (DSO) due to the seroconversion window, and for ENZY-WELL SARS-CoV-2 IgG and IgA ELISAs resulted 100% in those samples collected after 10 and 12 DSO, respectively. The results showed that ENZY-WELL SARS-CoV-2 ELISAs may represent a valid option for both diagnostic and epidemiological purposes, covering all different antibody types developed in SARS-CoV-2 immune response.

2011 ◽  
Vol 35 (1) ◽  
pp. 16-24
Author(s):  
Lazem H. Al-Taie

Back ground: Toxoplasmosis is an important zoonosis that causes economic losses in animal herds due to abortion and stillbirth as well as changes in the reproductive and neural system of susceptible animals . Objective: The aims of the present study is to determination the prevalence of T. gondii in farm animals ( sheep& goat)of both genders and different ages in Sulaimani province by using two serological tests (ELISA and LAT). Methods: Blood samples were collected from farm animals ,142 sheep and 46 goats , of different sexes and ages. Tow different serological tests ,ELISA and LAT for qualitative determination of T. gondii antibody titer in sheep and goats serum samples. Results: The prevalence rate in sheep was 73 (51.7 %) and 82 (57 %) , and 21 (54.6 %) and 25 (54.35 %) in goats ,by ELISA and LAT respectively. The prevalence of toxoplasmosis was highest in age group 7-9 (66.6%) in sheep in compares’ with other age groups. There was no significant differences between both spp.and tow test. Conclusion: Statistical results show no significant differences between both tests (ELISA &LAT) at (P ≥ 0.05).The prevalence of toxoplasmosis was increased proportionally with the age of animals, while gender has no effect on the prevalent rate .


2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Laura Sponton ◽  
Hulin Jin ◽  
Markus Fluck ◽  
Yusuke Suzuki ◽  
Amy Kao

Abstract Background and Aims Analysis of serum or plasma from patients with IgA nephropathy (IgAN) has confirmed the presence of elevated levels of circulating immune complexes containing Gd-IgA1 (Czerkinsky 1986). New sensitive and reasonably specific noninvasive tests are emerging to guide the therapeutic strategy that is applicable to all stages of IgAN (Suzuki 2014). Here we are reporting the fit for purpose validation of an ELISA method for the quantitative determination of Gd-IgA1 in human serum samples to support biomarker investigations in clinical studies of Merck KGaA, Darmstadt. Method The assay was developed based on a commercially available immunoassay kit. The dynamic range of the calibration curve was determined from 1.56 ng/mL (LLOQ) to 100 ng/mL (ULOQ). With a minimum required dilution of 200-fold and standard assay volume of 50.0 μL, the range of the method in matrix was from 312 ng/mL to 20, 000 ng/mL. In assay validation phase, multiple validation parameters were evaluated, which included minimum required dilution (MRD), calibration curve, matrix effect, Intra- & Inter run accuracy & precision, selectivity, and parallelism. Additional validation parameters include sample stability (short/long term, freeze-thaw) and batch-to-batch comparison. Results All samples measured for intra & Inter - assay precision, accuracy, fulfilled the specifications according to the acceptance criteria. The selectivity was assessed using blank serum matrix from 10 individuals: the result indicated that matrix components in serum did not interfere with the detection of Gd-IgA1. Parallelism assessment was performed successfully for both samples from healthy donors and IgAN patient samples up to dilution factor (DF) 3200 (serum samples from healthy donors were determined up to DF 1600). All DF-corrected results within the assay range were determined with %CV ≤ 30.0%. Batch to batch comparison was assessed successfully based on the known shelf life of the kit. Short term stability using QC samples were given for up to 24hrs at room temperature. Freeze-thaw stability was given for up to 3 cycles at -20°C±5°C and -75°C±15°C. The investigations were performed according to general guidelines for method validation and applicable regulations. The results of investigated validation parameters fulfilled the requirements and recommendations, generally accepted for bioanalytical projects. Conclusion The present validation qualified the method for the quantitative determination of Gd-IgA1 in human serum samples from clinical studies.


2011 ◽  
Vol 57 (6) ◽  
pp. 905-909 ◽  
Author(s):  
Kaname Ohyama ◽  
Yukitaka Ueki ◽  
Atsushi Kawakami ◽  
Naoya Kishikawa ◽  
Mami Tamai ◽  
...  

BACKGROUND Analysis of circulating immune complexes (CICs) produced during an immune response may be useful in elucidating some aspects of this process. Identification of antigens incorporated into CICs provides information that may be helpful in developing diagnostic and treatment strategies for autoimmune diseases, infection, cancer, and transplantation therapy, and such information might be more relevant than information on free antigens. Because CICs may contain many antigens, comprehensive identification and profiling of such antigens is more effective than immunoblotting detection. METHODS We developed a novel proteomic strategy (immune complexome analysis) in which immune complexes (ICs) are separated from serum, digested directly with trypsin, and then subjected to nano-liquid chromatography–tandem mass spectrometry for identifying and profiling antigens in CICs. We applied this strategy to the analysis of CICs in 21 rheumatoid arthritis (RA) patients. Serum samples from 13 healthy donors and 8 osteoarthritis patients were used as controls. RESULTS CICs containing thrombospondin-1 (TSP-1) and platelet factor 4 (PF4) were found in the serum of 81% and 52% of RA patients, respectively, and in none of the controls. CONCLUSIONS The ICs in the serum of a majority of the RA patients contained TSP-1 or PF4, and these ICs may have potential as alternative biomarkers. Our technique for immune complexome analysis uses routine clinical samples, simple protocols, and widely available equipment. This method may be generally applicable to the study of the relationship between CICs and certain diseases associated with the immune response in animals and humans.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Risheng Li ◽  
Xu Deng ◽  
Longfei Xia

Abstract Diabetes has become a universal epidemic in recent years. Herein, the monitoring of glucose in blood is of importance in clinical applications. In this work, PtNi alloy nanoparticles homogeneously dispersed on graphene (PtNi alloy-graphene) was synthesized as a highly effective electrode material for glucose detection. Based on the modified PtNi alloy-graphene/glass carbon (PtNi alloy-graphene/GC) electrode, it is found that the PtNi alloy-graphene/GC electrode exhibited excellent electrocatalytic performance on glucose oxidation. Furthermore, the results from amperometric current–time curve show a good linear range of 0.5–15 mM with the limit of detection of 16 uM (S/N = 3) and a high sensitivity of 24.03 uAmM−1 cm−2. On account of the good selectivity and durability, the modified electrode was successfully applied on glucose detection in blood serum samples.


2017 ◽  
Vol 2017 ◽  
pp. 1-6 ◽  
Author(s):  
Liping Wu ◽  
Yue Liu ◽  
Rong Huang ◽  
Huawen Zhao ◽  
Weiqun Shu

A rapid, simple, and novel method for folate receptor α (FRα) determination is reported here. A probe of gold nanoparticles (Au NPs) modified with anti-FRα antibody was synthesized under the optimized conditions first. The antibody-modified Au NPs would aggregate when FRα was added to the probe for the specific interaction between antibody and antigen, resulting in the enhancement of resonance Rayleigh scattering (RRS) intensity. There is a linear relationship between the change of RRS intensity (ΔIRRS) and the concentration of FRα, with the detecting range of 0.50–37.50 ng·mL−1 and the limit of determination of 0.05 ng·mL−1. The determination of FRα in serum samples was realized with the advantages of high selectivity, high sensitivity, and easy operation.


Parasitology ◽  
1989 ◽  
Vol 99 (S1) ◽  
pp. S5-S19 ◽  
Author(s):  
R. M. E. Parkhouse ◽  
L. J. S. Harrison

SUMMARYA thorough study of parasitic helminth antigens is a pre-requisite for control programmes based on accurate immunochemical diagnosis, protection by vaccination and perhaps immune modulation to diminish pathological sequelae. Studies should be directed at the identification of those stage- or age-specific surface, secreted and somatic antigens which are involved in the host-parasite interactions responsible for immunity and/or pathology. Current methods of diagnosis of parasitic infections often fail to detect low-level patent infections, which incurs the risk of having a reservoir capable of perpetuating infections. There is, then, an urgent requirement for accurate immunochemical diagnosis, to be used in association with, and for the evaluation of, drug treatment and vector elimination, in parasite control programmes. Given the high sensitivity of current immunoassay technology, the only bar to establishing the necessary immunological tests is the choice of suitably specific antigen/antibody systems. Assays designed to detect parasite products or antigens are a major priority, as they indicate current infection, whereas those which detect antibody only indicate exposure to infection, which may or may not be current. Surface and secreted antigens are the most likely targets for protective immune responses and thus form a logical focus for vaccine design. The cestodes, which present such strong evidence for immunity following natural infection, are likely to yield effective vaccines by modern procedures. Certain antigens must, however, stimulate the humoral and/or cellular responses which are responsible for the undesirable immunopathological consequences of many helminthic diseases. The nematodes and trematodes furnish some extreme examples of such pathology. The ultimate objective in identifying these particular antigens is to utilize them in the appropriate down-regulation of the immune response responsible for such pathology. As an illustration, we have presented an interesting correlation between one particular clinical condition of onchocerciasis (Sowda) and the serological response, defined both in terms of the parasite antigens and an immunoglobulin class-restricted antibody response. Finally, the complexity of these parasite systems and the host response to the parasite should not be underestimated. Modern analytical techniques allow their detailed analysis in terms of the humoral antibody responses and afford the possibility of the future development of control and disease management procedures tailored to each individual host-parasite system. However, novel systems are required to complete the analysis of the cellular components of the immune response to parasite antigens, and functional studies are needed to determine the role that these parasite antigens play in the complex interaction between parasite and host.


Polymers ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 2173
Author(s):  
Natalija German ◽  
Almira Ramanaviciene ◽  
Arunas Ramanavicius

Biosensors for the determination of glucose concentration have a great significance in clinical diagnosis, and in the food and pharmaceutics industries. In this research, short-chain polyaniline (PANI) and polypyrrole (Ppy)-based nanocomposites with glucose oxidase (GOx) and 6 nm diameter AuNPs (AuNPs(6 nm)) were deposited on the graphite rod (GR) electrode followed by the immobilization of GOx. Optimal conditions for the modification of GR electrodes by conducting polymer-based nanocomposites and GOx were elaborated. The electrodes were investigated by cyclic voltammetry and constant potential amperometry in the presence of the redox mediator phenazine methosulfate (PMS). The improved enzymatic biosensors based on GR/PANI-AuNPs(6 nm)-GOx/GOx and GR/Ppy-AuNPs(6 nm)-GOx/GOx electrodes were characterized by high sensitivity (65.4 and 55.4 μA mM−1 cm−2), low limit of detection (0.070 and 0.071 mmol L−1), wide linear range (up to 16.5 mmol L−1), good repeatability (RSD 4.67 and 5.89%), and appropriate stability (half-life period (τ1/2) was 22 and 17 days, respectively). The excellent anti-interference ability to ascorbic and uric acids and successful practical application for glucose determination in serum samples was presented for GR/PANI-AuNPs(6 nm)-GOx/GOx electrode.


2000 ◽  
Vol 7 (6) ◽  
pp. 964-966 ◽  
Author(s):  
Patricia Cordoba ◽  
Alejandra Lanoel ◽  
Sergio Grutadauria ◽  
Marta Zapata

ABSTRACT The protective immune responses against rubella virus (RV) are related to its neutralizing epitopes, an issue that is important to consider when assessing the immune status of patients with remote infection. In the present paper, we compare the antibodies detected by a synthetic-peptide-based enzyme immunoassay (EIA) with antibodies detected by the traditional technique of hemagglutination inhibition (HIA) in patients with remote RV infection. The synthetic peptide used as an antigen (SP15) represents a neutralizing epitope that corresponds to amino acids 208 to 239 of the E1 glycoprotein. The SP15-EIA was developed, all variables that affected the assay were standardized, and the test was validated using reference sera. Serum samples (n = 129) from patients with remote RV infection were tested by HIA and SP15-EIA. Discrepant sera were assayed by MEIA (IMX/Abbot). The comparison between HIA and SP15-EIA, taking HIA as the standard methodology for determining immune status, showed that SP15-EIA is very specific and sensitive for detecting protecting antibodies (specificity, 100%; sensitivity, 98.20%). This study demonstrates that antibodies against the neutralizing domain represented by SP15 would be important in the memory response after natural infection and may be a good tool in the determination of the true immune status of patients with remote infection with regard to RV.


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