An Enzyme-Linked Immunosorbent Assay for Detection of Bordetella avium Infection in Turkey Flocks: Sensitivity, Specificity, and Reproducibility

1991 ◽  
Vol 35 (2) ◽  
pp. 308 ◽  
Author(s):  
Elie K. Barbour ◽  
M. Kim Brinton ◽  
Susan D. Torkelson ◽  
James B. Johnson ◽  
Peter E. Poss
2007 ◽  
Vol 14 (3) ◽  
pp. 318-319 ◽  
Author(s):  
Ana Laín ◽  
María D. Moragues ◽  
Juan Carlos García Ruiz ◽  
Joaquín Mendoza ◽  
Ana Camacho ◽  
...  

ABSTRACT The performance of a new test to detect antibodies to Candida albicans recombinant enolase was investigated in 47 immunocompromised and 51 immunocompetent patients. The sensitivity, specificity, and positive and negative predictive values of the test for the diagnosis of invasive candidiasis were 81.0, 83.9, 79.1, and 85.5%, respectively.


2016 ◽  
Vol 23 (3) ◽  
pp. 282-286 ◽  
Author(s):  
Brianne M. Ritchie ◽  
Jean M. Connors ◽  
Katelyn W. Sylvester

Background: Previous studies have demonstrated optimized diagnostic accuracy in utilizing higher antiheparin–platelet factor 4 (PF4) enzyme-linked immunosorbent assay (ELISA) optical density (OD) thresholds for diagnosing heparin-induced thrombocytopenia (HIT). We describe the incidence of positive serotonin release assay (SRA) results, as well as performance characteristics, for antiheparin–PF4 ELISA thresholds ≥0.4, ≥0.8, and ≥1.0 OD units in the diagnosis of HIT at our institution. Methods: Following institutional review board approval, we conducted a single-center retrospective chart review on adult inpatients with a differential diagnosis of HIT evaluated by both antiheparin–PF4 ELISA and SRA from 2012 to 2014. The major endpoints were to assess incidence of positive SRA results, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy at antiheparin–PF4 ELISA values ≥0.4 OD units when compared to values ≥0.8 and ≥1.0 OD units. Clinical characteristics, including demographics, laboratory values, clinical and safety outcomes, length of stay, and mortality, were collected. Results: A total of 140 patients with 140 antiheparin–PF4 ELISA and SRA values were evaluated, of which 23 patients were SRA positive (16.4%) and 117 patients were SRA negative (83.6%). We identified a sensitivity of 91.3% versus 82.6% and 73.9%, specificity of 61.5% versus 87.2% and 91.5%, PPV of 31.8% versus 55.9% and 63.0%, NPV of 97.3% versus 96.2% and 94.7%, and accuracy of 66.4% versus 86.4% and 88.6% at antiheparin–PF4 ELISA thresholds ≥0.4, ≥0.8, and ≥1.0 OD units, respectively. Conclusion: Our study suggests an increased antiheparin–PF4 ELISA threshold of 0.8 or 1.0 OD units enhances specificity, PPV, and accuracy while maintaining NPV with decreased sensitivity.


1988 ◽  
Vol 32 (2) ◽  
pp. 353 ◽  
Author(s):  
B. A. Hopkins ◽  
J. K. Skeeles ◽  
G. E. Houghten ◽  
J. D. Story

1996 ◽  
Vol 40 (3) ◽  
pp. 637-641 ◽  
Author(s):  
J D Cleary ◽  
S W Chapman ◽  
J Deng ◽  
C J Lobb

Our purpose was to develop and characterize an enzyme-linked immunosorbent assay (ELISA) which could measure the concentration of amphotericin B in serum. Amphotericin B was assayed by competition ELISA. Multiwell ELISA plates coated with amphotericin B (1.0 micrograms/ml) conjugated to bovine serum albumin were used to test replicates of serum samples spiked with amphotericin B. Purified rabbit polyclonal antibody against amphotericin B (1.4 micrograms/ml) was added subsequent to the instillation of samples spiked with unknown amounts of amphotericin B. Experiments were performed to test the sensitivity, specificity, precision, and accuracy of the assay. The ability to measure lipid-associated amphotericin B was also evaluated in preliminary studies. Analysis of reference samples containing amphotericin B yielded a traditional sigmoidal curve. The limits of detection were 0.15 to 156 micrograms/ml. The sensitivity of the assay was affected by light and temperature exposure. Assay specificity was altered only by the presence of nystatin, a polyene antifungal agent similar to amphotericin B. Intrarun (coefficient of variation = 3.0%) and interrun (coefficient of variation = 12.8%) coefficients of variation were calculated and were comparable to those in similar assays. The assay's correlation coefficient (r = 0.907) demonstrated a statistically significant correlation between the optical density of the sample and the concentration of drug in the sample. The amphotericin B ELISA's ease, precision, and overall accuracy suggest that this assay could be used for assessments of serum amphotericin B concentrations. Multiple research questions concerning the role of serum amphotericin B concentrations in toxicity and efficacy have gone unanswered because of the labor-intensive nature of the assays which have been available to date. The ability to easily and rapidly measure 40 duplicate samples containing amphotericin B should also prove to be a distinct advantage for clinical research or reference laboratories in addressing these questions.


1987 ◽  
Vol 33 (6) ◽  
pp. 761-764 ◽  
Author(s):  
M Maeda ◽  
H Arakawa ◽  
A Tsuji ◽  
Y Yamagami ◽  
A Isozaki ◽  
...  

Abstract In this rapid, cost-effective, enzyme-linked immunosorbent assay for 17 alpha-hydroxyprogesterone (17-OHP) eluted from dried blood spotted on filter paper, second antibody is coated onto the microwell plate and horseradish peroxidase (EC 1.11.1.7) is the label enzyme. Antiserum to 17-OHP was prepared by using 4-(2-carboxymethylthio)-17-OHP-bovine serum albumin conjugate as immunogen. Enzyme conjugate was prepared from 4-(2-carboxymethylthio)-17-OHP and peroxidase. The blood spots are assayed in the microwells without extraction or centrifugation steps. The detection limit of the assay is 1 microgram/L, equivalent to 3.5 pg (10.6 fmol) per disc. Intra- and interassay CVs at two steroid concentrations (7.38 and 22.79 micrograms/L) ranged from 3.74 to 11.90% (n = 5), and 9.49 and 9.83% (n = 5), respectively. Results correlated well (r = 0.91) with those of a fluorescence enzyme immunoassay. The sensitivity, specificity, and precision of this method make it potentially useful in the mass screening of neonates for congenital adrenal hyperplasia.


2016 ◽  
Vol 4 (1) ◽  
pp. 178-184
Author(s):  
Rajesh Sharma ◽  
Pankaj Sharma ◽  
Gaush Talat ◽  
Praveen Gautam ◽  
Reba Chhabra ◽  
...  

The publication deals with a brief overview of Hepatitis C Virus (HCV) and donor blood screening for HCV by using conventional Rapid, Enzyme Linked Immunosorbent Assay (ELISA) and Chemiluminescence Immunoassay (CLIA) also. The advantages of various generation of HCV tests in terms of sensitivity, specificity and reduction in window period are discussed.


2005 ◽  
Vol 12 (10) ◽  
pp. 1152-1156 ◽  
Author(s):  
Chaisiri Wongkham ◽  
Chairat Tantrawatpan ◽  
Pewpan M. Intapan ◽  
Wanchai Maleewong ◽  
Sopit Wongkham ◽  
...  

ABSTRACT A cystatin capture enzyme-linked immunosorbent assay (ELISA) using recombinant Fasciola gigantica cathepsin L1 antigen was developed to detect specific immunoglobulin G (IgG) subclass antibodies (IgG1, IgG2, IgG3, and IgG4) and was evaluated for its diagnostic potential for human fasciolosis. In an analysis of the sera of 13 patients infected with F. gigantica, 209 patients with other parasitic infections, 32 cholangiocarcinoma patients, and 42 healthy controls, the IgG4-ELISA gave the highest diagnostic values. The sensitivity, specificity, accuracy, and positive and negative predictive values of this method based on the detection of IgG4 antibody were 100%, 99.3%, 99.3%, 86.7%, and 100%, respectively. The results revealed that restricting the ELISA to the detection of specific IgG4 antibody enhanced the specificity and accuracy for the serodiagnosis of human fasciolosis.


1988 ◽  
Vol 34 (9) ◽  
pp. 1767-1771 ◽  
Author(s):  
R Saïle ◽  
C Delpierre ◽  
P Puchois ◽  
G Hocke ◽  
C Cachera ◽  
...  

Abstract We describe a noncompetitive enzyme-linked immunosorbent assay for amyloid A apolipoprotein in human serum (apo SAA) in which specific antibodies against synthetic peptides are used. Microtiter plates were used as solid phase and coated with affinity-purified antibodies raised against SAA1-(95-104) peptide. After incubation with delipidated plasmas, the bound apo SAA was revealed by labeled antibodies raised against SAA1-(58-69) peptide. The assay offers several advantages over existing techniques: sensitivity, specificity, simplicity, and non-use of radioisotopes. Results correlate well with those by a nephelometric method in which polyclonal antibodies are used.


Author(s):  
H Miller ◽  
P Simpson ◽  
M Forman ◽  
A Prigan ◽  
S Kehl ◽  
...  

Infection with human cytomegalovirus (CMV) is common and may have grave consequences in transplant recipients and congenitally infected children. Diagnosis of CMV infection is based on detection of specific antibodies and molecular assays. The incorporation of CMV serological assays into diagnostic algorithms requires careful evaluation and interpretation. Very few serological assays measure CMV infection by a specific strain. We developed an enzyme-linked immunosorbent assay (ELISA) using CMV-encoded UL144 as antigen. UL144 encodes for three major genotypes, A, B and C, and recombinants. The ELISA was developed with the three UL144 proteins and optimized as a multiplex assay. Sera from 55 positive and 59 negative CMV IgG, determined by the clinical microbiology laboratory, were used for evaluation and optimization. A cutoff optical density (OD) that distinguishes UL144 antibody-positive from -negative sera was established. Sera detected UL144 A, B, C and combination of these antigens. Assay threshold of 0.1 was established, and from a total of 303 sera, the overall sensitivity, specificity, positive and negative predictive values of the multiplex ELISA were 86.72% (95% CI 79.59% to 92.07%), 96.57% (92.69% to 98.73%), 94.40% (88.45% to 97.38%), and 91.60% (87.50% to 94.44%). The inter- and intraassay median coefficients of variation were 0.06 (IQR 0.56, 0.2) and 0.171 (IQR 0.038, 0.302). No cross reactivity was observed with HSV-positive CMV-negative sera. This ELISA gives simple and reproducible results for detection of anti-CMV UL144 IgG. It may assist in differentiating natural infection from CMV vaccines that lack UL144, and may provide an important tool for epidemiological studies of CMV strains.


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