Accuracy Of Enzyme-linked Immunosorbent Assay Research Articles

Clinical pitfalls and serological diagnostics of MuSK myasthenia gravis

BackgroundWe aimed to evaluate the diagnostic accuracy of enzyme-linked immunosorbent assay (ELISA) for anti-muscle specific tyrosine kinase (MuSK) antibody (Ab) in a large cohort of anti-acetylcholine receptor (AChR) Ab-negative generalized myasthenia gravis (MG), and also to investigate clinical contexts for the diagnosis of MuSK MG.MethodsA retrospective study of 160 patients with a clinical suspicion of AChR Ab-negative generalized MG was performed. The serum samples were tested for anti-clustered AChR Ab by cell-based assay (CBA), anti-MuSK Ab by ELISA, CBA and/or radioimmunoprecipitation assay (RIPA). Clinical data were compared between anti-MuSK Ab-positive MG and double seronegative (AChR and MuSK) MG groups.ResultsAfter excluding non-MG and clustered AChR Ab-positive patients, we identified 89 patients as a cohort of AChR Ab-negative generalized MG. Anti-MuSK Ab was positive by ELISA in 22 (24.7%) patients. While CBA identified five additional anti-MuSK Ab-positive patients, the results of ELISA were mostly consistent with CBA and RIPA with Cohen’s kappa of 0.80 and 0.90, respectively (p < 0.001). The most frequent differential diagnosis was motor neuron disease particularly of bulbar onset which showed remarkably overlapping clinical and electrophysiological features with MuSK MG at presentation.ConclusionWhile confirming the highest sensitivity of CBA for detecting anti-MuSK Ab, our results highlight the clinical pitfalls in making a diagnosis of MuSK MG and may support a diagnostic utility of MuSK-ELISA in clinical practice.

The Accuracy of Enzyme-Linked Immunosorbent Assay in the Diagnosis of COVID-19 in Iranian Children

Background The present study aimed to evaluate the accuracy of enzyme-linked immunosorbent assay (ELISA) compared to the real-time reverse transcription-polymerase chain reaction (rRT-PCR) in the diagnosis of coronavirus disease 2019 (COVID-19) in Iranian children. Materials and Methods In this cross-sectional study, 90 children under 15 years of age were randomly selected from suspects of COVID-19 referred to the Tabriz Children Hospital, the main pediatric COVID-19 diagnostic center of Tabriz, from May 21, 2020 to June 21, 2020. Blood and nasopharyngeal samples were taken simultaneously at the referring time. The diagnostic accuracy of ELISA-based IgM and IgG antibody tests for COVID-19 were compared with the rRT-PCR. Results The calculated sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio, negative likelihood ratio, overall diagnostic accuracy, and diagnostic odds ratio were 0.5745, 0.9767, 0.9643, 0.6774, 24.66, 0.4357, 0.7667, and 56.60 for IgM; and 0.6170, 0.9302, 0.9355, 0.6897, 8.84, 0.4117, 0.7667, and 21.47 for IgG, respectively. Conclusion Due to the lower sensitivity of antibody detection-based serological tests compared to rRT-PCR, they cannot be considered as initial and reliable tests for the diagnosis of COVID-19. It can be suggested that the serological tests be only used as complementary tests to rRT-PCR or for monitoring the immune response of children with COVID-19.

ELISA versus Nested PCR for Detection of Hepatitis B Co-infection in HIV-infected People in Bandung Indonesia

The number of Human Immunodeficiency Virus infected patients in Bandung is increasing. HIV patients are often suffering from Hepatitis B Virusco-infection. The co-infection status must be detected using the accurate method. This study aims to compare the accuracy of Enzyme-Linked Immunosorbent Assay and nested Polymerase Chain Reaction in detecting HIV-HBV co-infection in Bandung. The research method used is experimental. The sample used in this study was 50 people. All samples had met the inclusion criteria. The presence of hepatitis B surface antigen in the serum was detected using a qualitative ELISA Wantai kit, while sHBsAg gene in the whole blood was amplified using 2 pairs of the specific primary. The results obtained from both methods were analyzed to determine sensitivity, specificity, and accuracy values. The result of Hepatitis B detection using ELISA method showed that 15 samples (30%) were positive, 1 sample (2%) was a false positive, and 3 samples (6%) were the false negative. Moreover, the nested PCR method showed that 18 samples (36%) were positive without false positive or false negative result. These results showed that ELISA has a sensitivity value of 83.33%, specificity of 96.87%, and the accuracy of 92%, while Nested PCR has a value of 100% for all three parameters. Therefore, it can be concluded that the Nested PCR method was more accurate than ELISA to detect Hepatitis B co-infection in HIV patients in Bandung.The Nested PCR can be recommended for detecting the other co-infection cases.&#x0D; &#x0D; Keywords: accuracy, HBsAg, co-infection, sensitivity, specificity

Diagnostic accuracy of Enzyme-Linked Immunosorbent Assays to detect anti-Leishmania antibodies in patients with American Tegumentary Leishmaniasis: a systematic review.

ABSTRACTAmerican Tegumentary leishmaniasis (ATL) is an infectious disease caused by several species of Leishmania . Even though the direct detection of parasites has low sensitivity, it is still the gold standard for the laboratory diagnosis of ATL. Recent studies have shown promising results of Enzyme-Linked Immunosorbent Assays ( ELISAs) using recombinant antigens. The aim of this study is to compare the accuracy of ELISAs using novel antigens with the standard ELISA based on soluble antigens of Leishmania (SLA) to diagnose ATL. Studies that analyzed patients with ATL and studies that evaluated the diagnostic accuracy of ELISAs using novel antigens and SLA were included. The Fourteen studies from PubMed, Regional Portal of the Virtual Health Library (BVS), Brazilian Society of Dermatology, Virtual Health Library (IBECS), Literature in the Health Sciences in Latin America and the Caribbean (LILACS), Medical Literature Analysis and Retrieval System Online (Medline), Elsevier Embase, Cochrane Library, The National Institute for Health and Care Excellence (NICE), and Cumulative Index to Nursing and Allied Health Literature (CINAHL) were included. The novel ELISA antigens showed a high sensitivity (93.8%-100%) and specificity (82.5-100%), a better diagnostic performance than SLA-based ELISAs (1-97.4% and 57.5-100%, respectively). Only 10 studies analyzed cross-reactions in serum samples from patients with Chagas disease, and only two studies reported a percentage of cross-reactivity. In this systematic review, the novel ELISA antigens showed better sensitivity and specificity with respect to SLA-based ELISAs. However, a meta-analysis should be performed to confirm this finding.

Accuracy of Enzyme-Linked Immunosorbent Assays (ELISAs) in Detecting Antibodies against Mycobacterium leprae in Leprosy Patients: A Systematic Review and Meta-Analysis.

IgM against Mycobacterium leprae may be detected by enzyme-linked immunosorbent assays (ELISAs) based on phenolic glycolipid I (PGL-I) or natural disaccharide octyl bovine serum albumin (ND-O-BSA) as antigens, and the IgG response can be detected by an ELISA based on lipid droplet protein 1 (LID-1). The titers of antibodies against these antigens vary with operational classification. The aim of this study was to compare the accuracy of ELISAs involving PGL-I and ND-O-BSA with that involving LID-1. We included studies that analyze multibacillary and paucibacillary leprosy cases and evaluate the diagnostic accuracy of ELISAs based on LID-1 and/or PGL-I or ND-O-BSA as antigens to measure antibody titers against M. leprae. Studies were found via PubMed, the Virtual Health Library Regional Portal, Literatura Latino-Americana e do Caribe em Ciências da Saúde, Índice Bibliográfico Espanhol de Ciências de Saúde, the Brazilian Society of Dermatology, National Institute for Health and Clinical Excellence, Cochrane Library, Embase (the Elsevier database), and Cumulative Index to Nursing and Allied Health Literature. The Quality Assessment of Diagnostic Accuracy Studies served as a methodological validity tool. Quantitative data were extracted using the Standards for Reporting of Diagnostic Accuracy. Sensitivity, specificity, and a diagnostic odds ratio were calculated, and a hierarchical summary receiver-operating characteristic curve and forest plots were constructed. The protocol register code for this meta-analysis is PROSPERO 2017: CRD42017055983. Nineteen studies were included. ND-O-BSA showed better overall performance in terms of sensitivity, specificity, positive and negative likelihood ratios, and diagnostic odds ratio when compared with PGL-I and LID-1. The multibacillary group showed better performance on these parameters (than the paucibacillary group did), at 94%, 99%, 129, 0.05, and 2293, respectively. LID-1 did not provide any advantage regarding the overall estimate of sensitivity in comparison with PGL-I or ND-O-BSA.

Diagnostic accuracy of enzyme-linked immunosorbent assay (ELISA) and immunoblot (IB) for the detection of antibodies against Neospora caninum in milk from dairy cows.

The aim of the study was to estimate the sensitivity and specificity of enzyme-linked immunosorbent assay (ELISA) and immunoblot (IB) for detecting antibodies of Neospora caninum in dairy cows, in the absence of a gold standard. The study complies with STRADAS-paratuberculosis guidelines for reporting the accuracy of the test. We tried to apply Bayesian models that do not require conditional independence of the tests under evaluation, but as convergence problems appeared, we used Bayesian methodology, that does not assume conditional dependence of the tests. Informative prior probability distributions were constructed, based on scientific inputs regarding sensitivity and specificity of the IB test and the prevalence of disease in the studied populations. IB sensitivity and specificity were estimated to be 98.8% and 91.3%, respectively, while the respective estimates for ELISA were 60% and 96.7%. A sensitivity analysis, where modified prior probability distributions concerning IB diagnostic accuracy applied, showed a limited effect in posterior assessments. We concluded that ELISA can be used to screen the bulk milk and secondly, IB can be used whenever needed.

Accuracy of enzyme-linked immunosorbent assays for quantification of antibodies against Aleutian mink disease virus

There is a growing interest among mink ranchers to select their stock for tolerance to the Aleutian mink disease virus (AMDV). Enzyme-linked immunosorbent assays (ELISA) are used to identify mink which have low anti-AMDV antibody titres and are expected to tolerate the AMDV infection. The objective of this study was to calculate the accuracy of three ELISA systems which were performed on blood or serum of AMDV-inoculated American mink (Neovison vison) at five laboratories in Canada, USA, Finland, the Netherlands and Denmark. The accuracy was determined by comparing the ELISA results with antibody titres measured by the counter-immunoelectrophoresis (CIEP) using 10 two-fold serial dilutions of the plasma. Antibody titres of 880 black mink which were inoculated with a spleen homogenate from a naturally infected mink were measured between 16 and 176 weeks post-inoculation. Each ELISA result from every laboratory covered a wide range of antibody titres and the Spearman’s rank correlation coefficients between CIEP and ELISA results from different laboratories varied between 0.41 and 0.83, indicating a low to moderate accuracy of ELISA systems for ranking mink by antibody titre. The recombinant VP2-based ELISA used in the Netherlands and Finland ranked the mink by antibody titres more accurately than did the AMDV-G-based ELISA platforms developed in Denmark and the USA, suggesting that the source of antigen was one of the factors affecting the accuracy of ELISA results. It was concluded that the ELISA systems, particularly those based on AMDV-G antigen, require further refinement to improve their accuracy for ranking mink by antibody titre.

Accuracy of Enzyme-Linked Immunosorbent Assay Using Crude and Purified Antigens for Serodiagnosis of Melioidosis

Five enzyme-linked immunosorbent assays developed to detect antibodies to different Burkholderia pseudomallei antigen preparations were evaluated as diagnostic tests for melioidosis in northeast Thailand. The highest diagnostic indices were observed for an affinity-purified antigen (sensitivity, 82%; specificity, 72%) and crude B. pseudomallei antigen (sensitivity, 81%; specificity, 70%), an improvement over the indirect hemagglutination assay (sensitivity, 73%; specificity, 64%).

Performance of Antibodies against Tissue Transglutaminase for the Diagnosis of Celiac Disease: Meta-Analysis

A meta-analysis of studies investigating the diagnostic accuracy of enzyme-linked immunosorbent assays (ELISA) for antibodies against tissue transglutaminases (tTG) of various origins in celiac disease (CD) diagnosis was carried out. Twenty-one studies, with untreated CD patients and healthy/CD-free controls, were included in the meta-analysis. The diagnostic accuracy was estimated using a summary receiver operating characteristic (SROC) curve and pooled sensitivity (Se) and specificity (Sp). Multiple assays within a study were treated by considering all the assays within a study and by analyzing the most popular assay (i.e., the commercial anti-tTTG ELISA most frequently utilized in the papers in which multiple assays were included). The SROC curve indicated the absence of heterogeneity, and the superiority of recombinant human tTG (rh-tTG) and purified human tTG (ph-tTG) compared to guinea pig-tTG (gp-tTG). The sensitivities (most popular assay) for rh-tTG, ph-tTG, and gp-tTG were 94%, 90%, and 92%, respectively, and the specificities were 97%, 92%, and 96%, respectively. A sensitivity analysis (exclusion of studies with bias) altered the results of ph-tTG: Se, 95%; Sp, 98%. The sensitivities (all individual assays) for rh-tTG, ph-tTG, and gp-tTG were 94%, 94%, and 91%, respectively, and the specificities were 95%, 94%, and 89%, respectively. Human tTG ELISA is sensitive and specific, and it can be used for mass screening. Sensitivity analysis showed that ph-tTG might perform better.

Validation of Accuracy of Enzyme-Linked Immunosorbent Assay in Hybridoma Screening and Proposal of an Improved Screening Method

The 96-well plate format of enzyme-linked immunosorbent assay (ELISA) is the de facto standard in screening hybridomas for active antibody. Despite its widespread use, there have been few or no systematic attempts to validate its accuracy and answer the fundamental question, is it finding all the positives? We report here on a comparison between ELISA and a semiautomated flow-based kinetic exclusion assay (KinExA), both used in screening the same hybridoma cell line. Our finding is that ELISA is both overreporting (false positives) and underreporting (false negatives) compared to the KinExA system. The large number of hybridoma cells (e.g., cultured in six 96-well plates) that must be checked is daunting in considering any method other than ELISA for routine screening. To overcome this, we devised a sampling strategy in which wells are combined in a specified pattern, allowing a significant reduction in the total number of measurements required.

The accuracy of enzyme-linked immunosorbent assay and latex agglutination progesterone test for the validation of estrus and early pregnancy diagnosis in dairy cattle

The accuracy of the enzyme-linked immunosorbent assay (ELISA) and the latex agglutination (LA) on-farm progesterone kit for detecting estrus and diagnosing early pregnancy was investigated in this study. Italian Friesian dairy cows (n=82) from 6 dairy herds were used for the collection of foremilk samples at the time of breeding and at 19, 21, and 23 days post insemination. Pregnancy status was ascertained by uterine palpation per rectum 40 to 60 days post insemination. Progesterone levels were affected by herd, percentage of milk fat, and the day of testing × diagnosis interaction. Validation of estrus by qualitative on-farm tests was 74.6% (LA) and 100.0% (ELISA) accurate using 0.5 ng/ml of progesterone as the RIA estimate for estrus. The accuracy rate for early pregnancy diagnosis by RIA was 68.4 to 83.8% for day 19 and day 21, respectively, while the detection rate for nonpregnancy was 84.6 to 100% on day 19 and day 21, respectively, as compared with uterine palpation per rectum. The average accuracy rate for early pregnancy diagnosis ranged from 84.7 to 92.3% for the LA and ELISA tests, respectively; the nonpregnancy rate was correctly predicted 93.9% to 68.2% for the LA and ELISA tests, respectively.