Improved Sensitivity and Specificity of Sandwich, Competitive and Capture Enzyme-Linked Immunosorbent Assays for Allergen-Specific Antibodies

1985 ◽  
Vol 77 (1-2) ◽  
pp. 198-200 ◽  
Author(s):  
D.M. Kemeny ◽  
R. Urbanek ◽  
D. Samuel ◽  
D. Richards
2013 ◽  
Vol 20 (3) ◽  
pp. 363-368 ◽  
Author(s):  
Jérôme T. J. Nicol ◽  
Rémy Robinot ◽  
Audrey Carpentier ◽  
Giovanni Carandina ◽  
Elisa Mazzoni ◽  
...  

ABSTRACTSix new human polyomaviruses have been identified since 2008 (Merkel cell polyomavirus [MCPyV], human polyomavirus 6 [HPyV6], HPyV7, HPyV9, trichodysplasia spinulosa polyomavirus [TSPyV], and Malawi polyomavirus [MWPyV]). The presence of specific antibodies against MCPyV, HPyV6, HPyV7, HPyV9, and TSPyV in 828 Italian subjects aged 1 to 100 years was investigated by virus-like particle-based enzyme-linked immunosorbent assays (ELISAs). The findings indicate that all of these new polyomaviruses circulate widely in humans, with seroprevalences in adulthood ranging from 39.4% for HPyV9 to 87.1% for MCPyV, and that primary exposure is most intense in childhood, with the exception of HPyV7 and HPyV9, for which the seroprevalence increased throughout life. The proportion of subjects with high antibody titers was found to increase with age for MCPyV and to decrease with age for TSPyV.


2006 ◽  
Vol 13 (5) ◽  
pp. 553-555 ◽  
Author(s):  
Xiaohong Huang ◽  
Xuenan Xuan ◽  
Rodolfo A. Verdida ◽  
Shoufa Zhang ◽  
Naoaki Yokoyama ◽  
...  

ABSTRACT An immunochromatographic test for the simultaneous detection of Babesia caballi- and B. equi-specific antibodies (BceICT) was developed using a recombinant B. caballi 48-kDa rhoptry protein (rBc48) and a recombinant truncated B. equi merozoite antigen 2 (rEMA-2t). An evaluation of the ability of the BceICT to detect antibodies in sera from uninfected horses and experimentally infected horses showed high sensitivities and specificities of 83.3% (10/12 sera) and 92.9% (52/56 sera), respectively, for the anti-B. caballi antibody and 94.1% (16/17 sera) and 88.2% (45/51 sera), respectively, for the anti-B. equi antibody. Results from the detection of antibodies in field-collected sera indicated that the BceICT results corresponded with those of enzyme-linked immunosorbent assays (ELISA), showing 91.8% correspondence (67/73 sera) for B. caballi and 95.9% correspondence (70/73 sera) for B. equi, and that the BceICT results also corresponded with the ICT for B. caballi and for B. equi, both of which were 98.2% (55/56 sera). The comparable results of the ICT and ELISA and the simplicity and rapidity of the performance of the ICT suggest that the BceICT would be a feasible test for the simultaneous serodiagnosis of both agents of equine babesiosis in the field.


Author(s):  
Sarah Hicks ◽  
Kai Pohl ◽  
Teresa Neeman ◽  
Hayley McNamara ◽  
Kate Parsons ◽  
...  

Estimates of seroprevalence of SARS-CoV-2 antibodies have been hampered by inadequate assay sensitivity and specificity. Using an ELISA-based approach to that combines data about IgG responses to both the Nucleocapsid and Spike-receptor binding domain antigens, we show that near-optimal sensitivity and specificity can be achieved. We used this assay to assess the frequency of virus-specific antibodies in a cohort of elective surgery patients in Australia and estimated seroprevalence in Australia to be 0.28% (0 to 0.72%). These data confirm the low level of transmission of SARS-CoV-2 in Australia before July 2020 and validate the specificity of our assay.


2009 ◽  
Vol 63 (1-2) ◽  
pp. 37-44
Author(s):  
Nenad Milic ◽  
Jakov Nisavic ◽  
Marina Radojicic ◽  
Marina Sekler ◽  
Kazimir Matovic ◽  
...  

Comparative investigations of the sensitivity and specificity of the indirect immunoenzyme probe - iELISA and the hemagglutination inhibition test (HI test) in serological diagnostics of the Newcastle disease in poultry were carried out using samples of blood serum taken from non-vaccinated and vaccinated poultry. A total of 14 samples of blood serum from non-vaccinated poultry were examined using the immunoenzyme probe - iELISA, and nine of these were found to be positive to the presence of specific antigen against the Newcastle disease virus, while two samples were suspect, and no presence of specific antibodies was established in three samples. Examinations of 82 samples of blood serum from vaccinated poultry for the presence of specific antibodies against the Newcastle disease virus established their presence in 80 serum samples, while one sample was suspect and one sample was negative. The values of the titer of specific antibodies in blood serum samples of vaccinated and non-vaccinated poultry established using the hemagglutination inhibition test (HI test) ranged from 1:2 to 1:32.


1990 ◽  
Vol 32 (2) ◽  
pp. 96-100 ◽  
Author(s):  
Jairo Ivo-Dos-Santos ◽  
Deise L. Campos Mello ◽  
José C. Couto-Fernandez ◽  
Roberto M. Passos ◽  
Leila A. Dias-Carneiro ◽  
...  

Sera from 472 Brazilian subjects, confirmed to be either positive or negative for HIV antibodies and comprising the total clinical spectrum of HIV infection, were utilized in the evaluation of six commercially available enzyme-linked immunosorbent assays (ELISA), as well as of four alternative assays, namely indirect immunofluorescence (IIF), passive hemagglutination (PHA), dot blot and Karpas AIDS cell test. The sensitivities ranged from 100% (Abbott and Roche ELISA) to 84.2% (PHA) and the specificities ranged from 99.3% (IIF) to 80.2% (PHA). The sensitivity and specificity of the PHA and the sensitivity of the Karpas cell test were significantly lower than those of the other tests. Although the IFF and dot blot had good sensitivities and specificities, the six ELISA were more attractive than those tests when other parameters such as ease of reading and duration of assay were considered.


2012 ◽  
Vol 19 (3) ◽  
pp. 334-337 ◽  
Author(s):  
Maying Tse ◽  
Mia Kim ◽  
Chung-Hei Chan ◽  
Po-Lai Ho ◽  
Siu-Kit Ma ◽  
...  

ABSTRACTThe reverse zoonotic transmission of the pandemic H1N1 2009 influenza virus to swine necessitates enhanced surveillance of swine for influenza virus infection. Using a well-characterized panel of naturally infected swine sera, we evaluated and optimized the performances of three commercially available competitive enzyme-linked immunosorbent assays (ELISAs), namely, the IDEXX Influenza A Ab test, IDEXX AI MultiS-Screen Ab test, and IDVet ID Screen influenza A antibody competition ELISA, for detecting influenza A virus-reactive antibodies in swine. Receiver operating characteristic (ROC) analysis suggests that adjustment of the manufacturer-recommended cutoff values optimizes the sensitivity and specificity of these assays, making them applicable for seroepidemiology studies of swine influenza. Using such optimized cutoff levels, the sensitivity and specificity of the IDEXX Influenza A Ab test were 86% and 89%, respectively; those for the IDEXX AI MultiS-Screen Ab test were 91% and 87%, respectively; and those for the IDVet ID Screen influenza A test were 95% and 79%, respectively.


1988 ◽  
Vol 100 (2) ◽  
pp. 205-212 ◽  
Author(s):  
R. A. Payne ◽  
D. H. M. Joynson ◽  
A. J. Wilsmore

SUMMARYTachyzoitcs of the RH strain of Toxoplasma gondii were inoculated intravenously into sheep following which serum samples were collected at approximately weekly intervals for 9 months. The sera were examined by the toxoplasma dye test and two enzymc-linkcd immunosorbent assays (ELISA) specifically developed for investigations of ovine toxoplasmosis. One was an antibody class capture assay for the detection of anti-toxoplasma specific IgM, the other an indirect assay which detected anti-toxoplasma IgG.Some of the sheep had antibodies to toxoplasma prior to inoculation but none had specific IgM. Sera collected 17 days after inoculation showed that all had raised specific antibody levels but the only sheep that produced specific antitoxoplasma IgM were those that were initially without any antibody. Specific IgM could be detected in all these particular sheep for at least 1 month after infection and up to 3 months in some. Specific IgG persisted at high levels for at least 3 months and could still be detected at moderate levels for at least 9 months. The ELISA methods described are simple to perform and could clearly distinguish between previous infection and this experimental infection with Toxoplasma gondii.


2020 ◽  
Vol 223 (1) ◽  
pp. 10-14 ◽  
Author(s):  
Sarah M Hicks ◽  
Kai Pohl ◽  
Teresa Neeman ◽  
Hayley A McNamara ◽  
Kate M Parsons ◽  
...  

Abstract Estimates of seroprevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies have been hampered by inadequate assay sensitivity and specificity. Using an enzyme-linked immunosorbent assay–based approach that combines data about immunoglobulin G responses to both the nucleocapsid and spike receptor binding domain antigens, we show that excellent sensitivity and specificity can be achieved. We used this assay to assess the frequency of virus-specific antibodies in a cohort of elective surgery patients in Australia and estimated seroprevalence in Australia to be 0.28% (95% Confidence Interval, 0–1.15%). These data confirm the low level of transmission of SARS-CoV-2 in Australia before July 2020 and validate the specificity of our assay.


Author(s):  
Steven E Conklin ◽  
Kathryn Martin ◽  
Yukari C Manabe ◽  
Haley A Schmidt ◽  
Morgan Keruly ◽  
...  

Background. Rapid point-of-care tests (POCTs) for SARS-CoV-2-specific antibodies vary in performance. A critical need exists to perform head-to-head comparison of these assays. Methods. Performance of fifteen different lateral flow POCTs for the detection of SARS-CoV-2-specific antibodies was performed on a well characterized set of 100 samples. Of these, 40 samples from known SARS-CoV-2-infected, convalescent individuals (average of 45 days post symptom onset) were used to assess sensitivity. Sixty samples from the pre-pandemic era (negative control), that were known to have been infected with other respiratory viruses (rhinoviruses A, B, C and/or coronavirus 229E, HKU1, NL63 OC43) were used to assess specificity. The timing of seroconversion was assessed on five POCTs on a panel of 272 longitudinal samples from 47 patients of known time since symptom onset. Results. For the assays that were evaluated, the sensitivity and specificity for any reactive band ranged from 55%-97% and 78%-100%, respectively. When assessing the performance of the IgM and the IgG bands alone, sensitivity and specificity ranged from 0%-88% and 80%-100% for IgM and 25%-95% and 90%-100% for IgG. Longitudinal testing revealed that median time post symptom onset to a positive result was 7 days (IQR 5.4, 9.8) for IgM and 8.2 days (IQR 6.3 to 11.3). Conclusion. The testing performance varied widely among POCTs with most variation related to the sensitivity of the assays. The IgM band was most likely to misclassify pre-pandemic samples. The appearance of IgM and IgG bands occurred almost simultaneously.


Author(s):  
Steven E. Conklin ◽  
Kathryn Martin ◽  
Yukari C Manabe ◽  
Haley A Schmidt ◽  
Jernelle Miller ◽  
...  

Background. Rapid point-of-care tests (POCTs) for SARS-CoV-2-specific antibodies vary in performance. A critical need exists to perform head-to-head comparison of these assays. Methods. Performance of fifteen different lateral flow POCTs for the detection of SARS-CoV-2-specific antibodies was performed on a well characterized set of 100 samples. Of these, 40 samples from known SARS-CoV-2-infected, convalescent individuals (average of 45 days post symptom onset) were used to assess sensitivity. Sixty samples from the pre-pandemic era (negative control), that were known to have been infected with other respiratory viruses (rhinoviruses A, B, C and/or coronavirus 229E, HKU1, NL63 OC43) were used to assess specificity. The timing of seroconversion was assessed on five LFAs on a panel of 272 longitudinal samples from 47 patients of known time since symptom onset. Results. For the assays that were evaluated, the sensitivity and specificity for any reactive band ranged from 55%-97% and 78%-100%, respectively. When assessing the performance of the IgM and the IgG bands alone, sensitivity and specificity ranged from 0%-88% and 80%-100% for IgM and 25%-95% and 90%-100% for IgG. Longitudinal testing revealed that median time post symptom onset to a positive result was 7 days (IQR 5.4, 9.8) for IgM and 8.2 days (IQR 6.3 to 11.3) for IgG. Conclusion. The testing performance varied widely among LFAs with most variation related to the sensitivity of the assays. The IgM band was most likely to misclassify pre-pandemic samples. The appearance of IgM and IgG bands occurred almost simultaneously.


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