scholarly journals Blastomyces dermatitidis Antigen Detection by Quantitative Enzyme Immunoassay

2011 ◽  
Vol 19 (1) ◽  
pp. 53-56 ◽  
Author(s):  
Patricia Connolly ◽  
Chadi A. Hage ◽  
J. Ryan Bariola ◽  
Eric Bensadoun ◽  
Mark Rodgers ◽  
...  
Keyword(s):  
Enzyme Immunoassay ◽  
Healthy Subjects ◽  
Antigen Detection ◽  
Antigen Level ◽  
Antigen Concentration ◽  
Content Type ◽  
Antigen Assay ◽  
Concurrent Testing ◽  
Urine And Serum ◽  
Urine Specimens

ABSTRACTThe second-generation MVistaBlastomycesantigen enzyme immunoassay was not quantitative; therefore, specimens obtained previously were tested in the same assay as new specimens to assess the change in antigen levels. Furthermore, the sensitivity in serum had not been fully evaluated. The purpose of this study was to evaluate a quantitativeBlastomycesantigen assay and detection of antigen in serum. Calibrators containing known concentrations ofBlastomycesgalactomannan were used to quantify antigen in urine and serum from patients with proven blastomycosis and from controls. Paired current and previously obtained urine specimens were tested to determine if quantification eliminated the need for concurrent testing to assess change in antigen. Pretreatment of serum with EDTA at 104°C was evaluated to determine if dissociation of immune complexes improved detection of antigenemia. Antigenuria was detected in 89.9% of patients with culture- or histopathology-proven blastomycosis. Specificity was 99.0% in patients with nonfungal infections and healthy subjects, but cross-reactions occurred in 95.6% of patients with histoplasmosis. Change in antigen level categorized as increase, no change, or decrease based on antigen units determined in the same assay agreed closely with the category of change in ng/ml determined from different assays. Pretreatment increased the sensitivity of detection of antigenemia from 35.7% to 57.1%. Quantification eliminated the need for concurrent testing of current and previously obtained specimens for assessment of changes in antigen concentration. Pretreatment increased the sensitivity for detection of antigenemia. Differentiation of histoplasmosis and blastomycosis is not possible by antigen detection.

scholarly journals Evaluation of Coccidioides Antigen Detection in Dogs with Coccidioidomycosis

2012 ◽  
Vol 19 (3) ◽  
pp. 343-345 ◽  
Author(s):  
Emily J. Kirsch ◽  
Russell T. Greene ◽  
Annalisa Prahl ◽  
Stanley I. Rubin ◽  
Jane E. Sykes ◽  
...  
Keyword(s):  
Enzyme Immunoassay ◽  
Fungal Infections ◽  
Rapid Diagnosis ◽  
Antigen Detection ◽  
Antibody Testing ◽  
Content Type ◽  
Healthy Dogs ◽  
Galactomannan Antigen ◽  
Antigen Antibody ◽  
Urine And Serum

ABSTRACTAntigen detection has been reported to be a promising method for rapid diagnosis of coccidioidomycosis in humans.Coccidioidesantigen detection has not been previously reported in dogs with coccidioidomycosis and was evaluated in 60 cases diagnosed based on detection of anti-Coccidioidesantibodies at titers of 1:16 or more in serum. Controls included dogs with presumed histoplasmosis or blastomycosis, other fungal infections, or nonfungal diseases and healthy dogs. Urine and serum specimens were tested using an enzyme immunoassay forCoccidioidesgalactomannan antigen. Antibody testing was performed at commercial veterinary reference laboratories. Antigen was detected in urine or serum of 12 of 60 (20.0%), urine only in 2 of 57 (3.5%), and serum only in 11 of 58 (19.0%) dogs with coccidioidomycosis. Antigen was detected in the urine of 3 of 43 (7.0%) and serum of 1 of 37 (2.7%) dogs with histoplasmosis or blastomycosis but not in 13 dogs with other fungal infections (serum, 9; urine, 13), 41 dogs with nonfungal diseases (urine, 41; serum, 18), or healthy dogs (serum, 21; urine, 21). Detection of antigen was an insensitive method for diagnosis of coccidioidomycosis in dogs in which the diagnosis was based primarily upon detection of antibodies at titers of 1:16 or higher, and the highest sensitivity was in serum.

scholarly journals Multiplex Urinary Antigen Detection for 13 Streptococcus pneumoniae Serotypes Improves Diagnosis of Pneumococcal Pneumonia in South African HIV-Infected Adults

2016 ◽  
Vol 55 (1) ◽  
pp. 302-312 ◽  
Author(s):  
Werner C. Albrich ◽  
Michael W. Pride ◽  
Shabir A. Madhi ◽  
Jan Callahan ◽  
Peter V. Adrian ◽  
...  
Keyword(s):  
South African ◽  
Pneumococcal Pneumonia ◽  
Antigen Detection ◽  
Urinary Antigen ◽  
Content Type ◽  
Urinary Antigen Detection ◽  
Dominant Serotype ◽  
Quellung Reaction ◽  
Urine Specimens ◽  
Asymptomatic Hiv

ABSTRACT A serotype-specific urinary antigen detection (UAD) assay for 13 serotypes included in the pneumococcal conjugate vaccine (PCV13) was recently reported as a useful diagnostic tool for pneumococcal pneumonia. We aimed to assess the diagnostic accuracy of the UAD in HIV-infected South African adults. Urine specimens from a well-defined cohort of HIV-infected South African adults with pneumonia were evaluated retrospectively in the UAD assay. Pneumonia was considered pneumococcal if either sputum Gram stain, sputum culture, blood culture, or the immunochromatographic (ICT) BinaxNow S. pneumoniae test (composite diagnostic) was positive. Among 235 enrolled pneumonia patients, the UAD assay was more frequently positive (104 [44.3%]) than the composite diagnostic (71 [30.2%]; P < 0.001) and increased the pneumococcal etiology from 30.2% by an additional 22.6% to 52.8%. The UAD assay detected more pneumococcal etiologies (45.0%) than the serotype-independent ICT (23.4%, P < 0.001). UAD identified 6/7 patients with PCV13 serotype bacteremia without misclassification of bacteremia episodes due to non-PCV13 serotypes. UAD was positive for 5.1% of asymptomatic HIV-infected persons, with higher rates among those with nasopharyngeal carriage. Concordance between serotypes identified by UAD and by Quellung reaction and PCR serotyping was 70/86 (81.4%). UAD identified the dominant serotype in multiple serotype carriage. This study confirms the utility of the UAD assay for HIV-infected adults comparing favorably with other diagnostic tests. A highly valent UAD may become a new standard for detection of pneumococcal pneumonia in adults. Prior to PCV introduction, at least 53% of pneumonia cases were due to pneumococci in HIV-infected South African adults.

scholarly journals Histoplasma Antigen Clearance during Treatment of Histoplasmosis in Patients with AIDS Determined by a Quantitative Antigen Enzyme Immunoassay

2011 ◽  
Vol 18 (4) ◽  
pp. 661-666 ◽  
Author(s):  
Chadi A. Hage ◽  
Emily J. Kirsch ◽  
Timothy E. Stump ◽  
Carol A. Kauffman ◽  
Mitchell Goldman ◽  
...  
Keyword(s):  
Enzyme Immunoassay ◽  
Amphotericin B ◽  
Latent Class ◽  
Response To Treatment ◽  
Separate Analysis ◽  
Content Type ◽  
Progressive Disseminated Histoplasmosis ◽  
Latent Class Growth ◽  
Kinetics Of ◽  
Urine And Serum

ABSTRACTClearance ofHistoplasmaantigen has been used as a marker for response to treatment of progressive disseminated histoplasmosis (PDH) in patients with AIDS. Advancements inHistoplasmaantigen detection permit accurate quantification of antigen concentration. We compared the clearance of antigenemia and antigenuria during effective treatment of PDH. Urine and serum specimens were serially collected from patients with AIDS who were successfully treated for PDH as part of two prospective clinical trials. Samples were stored frozen until they were tested in the quantitativeHistoplasmaantigen enzyme immunoassay. The kinetics of antigen clearance during the first 12 weeks of therapy were assessed in urine and serum during treatment with liposomal or deoxycholate amphotericin B followed by itraconazole and, in a separate analysis, in patients receiving only itraconazole. Latent class growth analysis was performed to define patterns of antigen clearance over time. In patients receiving amphotericin B, antigen levels declined the most during the first 2 weeks of treatment and antigenemia decreased more rapidly than antigenuria (5.90 ng/ml per week versus 4.21 ng/ml per week, respectively;P= 0.09). Mean reductions of antigen levels from baseline at weeks 2 and 12 were greater in sera than in urine: 11.26 ng/ml versus 7.65 ng/ml (P= 0.0948) and 18.52 ng/ml versus 14.64 ng/ml (P= 0.0440), respectively. In patients who received itraconazole alone, most of the decline in antigenuria occurred later during treatment and was overall slower than that seen with amphotericin B (P< 0.0001). Results of latent class growth modeling showed two distinct trajectories for each parameter. With effective therapy,Histoplasmaantigenemia decreases more rapidly than antigenuria, providing a more sensitive early laboratory marker for response to treatment. Antigenuria declines earlier with amphotericin B than with itraconazole.

scholarly journals Detection of Coccidioides Antigenemia following Dissociation of Immune Complexes

2009 ◽  
Vol 16 (10) ◽  
pp. 1453-1456 ◽  
Author(s):  
Michelle Durkin ◽  
Lois Estok ◽  
Duane Hospenthal ◽  
Nancy Crum-Cianflone ◽  
Samantha Swartzentruber ◽  
...  
Keyword(s):  
Heat Treatment ◽  
Enzyme Immunoassay ◽  
Immune Complexes ◽  
Healthy Subjects ◽  
Serum Samples ◽  
Cross Reactions ◽  
Heat Treated ◽  
Urine And Serum

ABSTRACT Having reported that pretreatment of serum samples with EDTA at 100°C improved the sensitivity for the detection of Histoplasma antigenemia, we have evaluated this method for the detection of Coccidioides antigenemia. Urine and serum samples from patients with coccidioidomycosis were tested using the MVista Coccidioides enzyme immunoassay, and serum samples with and without EDTA-heat treatment were tested. Antigenemia was detected in 28.6% of patients whose samples were not EDTA-heat treated and in 73.1% of those whose samples were treated. Antigenuria was detected in 50% of patients. Specificity of 100% was obtained in healthy subjects, but cross-reactions were seen in 22.2% of patients with histoplasmosis or blastomycosis. EDTA-heat treatment improves the sensitivity for the detection of Coccidioides antigenemia.

scholarly journals Advances in Diagnosis of Progressive Pulmonary and Disseminated Coccidioidomycosis

2020 ◽  
Author(s):  
Christelle Kassis ◽  
Michelle Durkin ◽  
Eric Holbrook ◽  
Robert Myers ◽  
Lawrence Wheat
Keyword(s):  
Cerebrospinal Fluid ◽  
Retrospective Study ◽  
Medical Center ◽  
Antigen Detection ◽  
Antibody Detection ◽  
Main Method ◽  
System Sensitivity ◽  
Serum Antigen ◽  
Urine And Serum ◽  
Immunocompetent Patients

Abstract Background Antibody detection is the main method for diagnosis of coccidioidomycosis, but it has limitations. The Coccidioides antigen enzyme immunoassay is recommended for testing cerebrospinal fluid in suspected meningitis. Reports on urine and serum antigen detection evaluated small numbers of patients who were mostly immunocompromised. The purpose of this study was to assess the accuracy of combined antibody and antigen detection for diagnosis. Methods A retrospective study, including all patients in whom Coccidioides antigen detection in serum was performed between January 2013 and May 2017, was conducted at Valleywise Health Medical Center (formerly Maricopa Integrated Health System). Sensitivity and specificity of antigen and antibody were evaluated in 158 cases and 487 controls. Results The sensitivity of antibody detection by immunodiffusion (ID) was 84.2%. The sensitivity of antigen detection was 57.0% if both urine and serum were tested and 36.7% if urine alone was tested. The sensitivity of combining antigen and ID antibody detection was 93.0%. The sensitivity of urine and serum antigen detection was 55.4% in proven and 58.7% in probable cases, 79.1% in disseminated and 41.6% in pulmonary cases, and 74.7% in immunocompromised and 40.0% in immunocompetent patients. Specificity was 99.4% for antigen detection and 96.5% for ID antibody detection. Diagnostic accuracy was 95.4% for ID antibody and antigen detection, 93.6% for ID antibody alone, and 89.1% for pathology or culture. Conclusions These findings support combined antibody and antigen detection for diagnosis of progressive coccidioidomycosis. The diagnosis may have been missed if antigen detection was not performed.

scholarly journals Culture of Urine Specimens by Use of chromID CPS Elite Medium Can Expedite Escherichia coli Identification and Reduce Hands-On Time in the Clinical Laboratory

2016 ◽  
Vol 54 (11) ◽  
pp. 2767-2773 ◽  
Author(s):  
Melanie L. Yarbrough ◽  
Meghan A. Wallace ◽  
Cynthia Marshall ◽  
Erin Mathias ◽  
Carey-Ann D. Burnham
Keyword(s):  
Escherichia Coli ◽  
Clinical Laboratory ◽  
Time Interval ◽  
Clinical Microbiology Laboratory ◽  
Biochemical Tests ◽  
Content Type ◽  
E Coli ◽  
Hands On ◽  
Clinically Significant ◽  
Urine Specimens

Urine is one of the most common specimen types submitted to the clinical microbiology laboratory; the use of chromogenic agar is one method by which the laboratory might expedite culture results and reduce hands-on time and materials required for urine culture analysis. The objective of our study was to compare chromID CPS Elite (bioMérieux), a chromogenic medium, to conventional primary culture medium for evaluation of urine specimens. Remnant urine specimens (n= 200) were inoculated into conventional media and into chromID CPS Elite agar (chromID). The time to identification and consumables used were documented for both methods. Clinically significant pathogen(s) were recovered from 51 cultures using conventional media, withEscherichia colibeing the most frequently recovered organism (n= 22). The rate of exact uropathogen agreement between conventional and chromogenic media was 82%, while overall categorical agreement was 83.5% The time interval between plating and final organism identification was decreased with chromID agar versus conventional media forE. coli(mean of 24.4 h versus 27.1 h,P< 0.001). Using chromID, clinically significant cultures required less hands-on time per culture (mean of 1 min and 2 s [1:02 min]) compared to conventional media (mean of 1:31 min). In addition, fewer consumables (2.4 versus 3.3 sticks and swabs) and rapid biochemical tests (1.0 versus 1.9) were necessary using chromID versus conventional media. Notably, antimicrobial susceptibility testing demonstrated good overall agreement (97.4%) between the chromID and conventional media for all antibiotics tested. chromID CPS Elite is accurate for uropathogen identification, reduces consumable usage, and may expedite the identification ofE. coliin clinical specimens.

Two-site enzyme immunoassay of CD4 and CD8 molecules on the surface of T lymphocytes from healthy subjects and HIV-1-infected patients

1994 ◽  
Vol 40 (1) ◽  
pp. 30-37 ◽  
Author(s):  
D Carriere ◽  
C Fontaine ◽  
A M Berthier ◽  
A M Rouquette ◽  
P Carayon ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
T Lymphocytes ◽  
Enzyme Immunoassay ◽  
Cd8 T Cells ◽  
Envelope Glycoprotein ◽  
Healthy Subjects ◽  
Gp 120 ◽  
Highly Sensitive ◽  
One Step ◽  
Hiv 1

Abstract A highly sensitive two-site enzyme immunoassay (Capcellia) was developed to determine the concentration of CD4 and CD8 molecules expressed on the surface of human T lymphocytes. This assay, performed in one step (20 min), involves the specific immunocapture of T lymphocytes and reaction of the CD4 or CD8 molecules with an enzyme-labeled monoclonal antibody (mAb). The results were expressed as molar concentrations of the T-cell markers on the basis of results obtained with calibrated CD4 and CD8 standards. The assay was sensitive enough to detect 0.4 pmol/L CD4 or 0.8 pmol/L CD8, which corresponded to approximately 20 x 10(6) CD4+ or CD8+ T cells per liter of blood. Mean concentrations in healthy adults were 17.2 pmol/L for CD4 and 22.1 pmol/L for CD8. The CD4 concentration was &lt; 8 pmol/L in 50% of HIV-1-infected patients and in 95% of AIDS patients. Given the epitopic specificity of the mAb to CD4 we used, these values correspond to the concentration of CD4 molecules free of envelope glycoprotein (gp)120.

A study of high-, middle- and low-molecular weight adiponectin in urine as a surrogate marker for early diabetic nephropathy using ultrasensitive immune complex transfer enzyme immunoassay

2018 ◽  
Vol 55 (5) ◽  
pp. 525-534 ◽  
Author(s):  
Mayumi Yamamoto ◽  
Yuki Fujimoto ◽  
Shino Hayashi ◽  
Seiichi Hashida
Keyword(s):  
Molecular Weight ◽  
Diabetic Nephropathy ◽  
Enzyme Immunoassay ◽  
Immune Complex ◽  
Healthy Subjects ◽  
Surrogate Marker ◽  
Urinary Albumin ◽  
Low Molecular Weight ◽  
Patients With Diabetes ◽  
Estimate Glomerular Filtration Rate

Background For the early identification of patients at risk of developing diabetic nephropathy, we have developed an ultrasensitive immune complex transfer enzyme immunoassay to measure adiponectin in urine. Methods We developed immune complex transfer enzyme immunoassay for adiponectin and measured urinary adiponectin from 70 healthy subjects, 35 obese non-diabetic subjects and 20 patients with diabetes. Results The urinary adiponectin concentrations in patients with diabetes (3.3 ± 10.7 ng/mg creatinine) were significantly higher than those in obese subjects (0.54 ± 0.44; P < 0.01) and healthy subjects (0.46 ± 0.42; P < 0.001). The gel filtration elution profile of urine from healthy subjects showed traces of four immunoreactive peaks (high-, medium-, low-molecular weight and monomer molecules), despite the majority of blood adiponectin being high-molecular weight. However, urinary adiponectin molecules were more frequent in low-molecular weight as the estimate glomerular filtration rate decreased. Furthermore, as blood glucose concentrations rose, middle-molecular weight and high-molecular weight increased in urine. Further, urinary adiponectin concentrations correlated with estimate glomerular filtration rate ( r = −0.61, P < 0.001), but not urinary albumin. In addition, our analysis showed a significantly ( P < 0.001) higher value for urinary adiponectin in the G2 stage of chronic kidney disease classification where urinary albumin is not elevated. Conclusion Adiponectin increases in urine as renal function decreases, and urinary adiponectin may be useful as a surrogate marker for diabetic nephropathy risk.

scholarly journals Flow Cytometry Analysis Using Sysmex UF-1000i Classifies Uropathogens Based on Bacterial, Leukocyte, and Erythrocyte Counts in Urine Specimens among Patients with Urinary Tract Infections

2014 ◽  
Vol 53 (2) ◽  
pp. 539-545 ◽  
Author(s):  
Tor Monsen ◽  
Patrik Rydén
Keyword(s):  
Flow Cytometry ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Flow Cytometry Analysis ◽  
Content Type ◽  
E Coli ◽  
New Information ◽  
Tract Infections ◽  
A Prospective Study ◽  
Urine Specimens

Urinary tract infections (UTIs) are the second most common bacterial infection. Urine culture is the gold standard for diagnosis, but new techniques, such as flow cytometry analysis (FCA), have been introduced. The aim of the present study was to evaluate FCA characteristics regarding bacteriuria, leukocyturia, and erythrocyturia in relation to cultured uropathogens in specimens from patients with a suspected UTI. We also wanted to evaluate whether the FCA characteristics can identify uropathogens prior to culture. From a prospective study, 1,587 consecutive urine specimens underwent FCA prior to culture during January and February 2012. Outpatients and inpatients (79.6% and 19.4%, respectively) were included, of whom women represented 67.5%. In total, 620 specimens yielded growth, of whichEscherichia colirepresented 65%,Enterococcusspp. 8%,Klebsiellaspp. 7%, andStaphylococcusspp. 5%. For the uropathogens, the outcome of FCA was compared against the results for specimens withE. coliand those with a negative culture.E. colihad high bacterial (median, 17,914/μl), leukocyte (median, 348/μl), and erythrocyte (median, 23/μl) counts. With the exception ofKlebsiellaspp., the majority of the uropathogens had considerable or significantly lower bacterial counts than that ofE. coli. High leukocyte counts were found in specimens withStaphylococcus aureus,Proteus mirabilis,Pseudomonas aeruginosa, and group C streptococci. Elevated erythrocyte counts were found forP. vulgaris,P. aeruginosa, and group C streptococci, as well as forStaphylococcus saprophyticus. In essence, FCA adds new information about the bacterial, leukocyte, and erythrocyte counts in urine specimens for different uropathogens. Based on FCA characteristics, uropathogens can be classified and identified prior to culture.E. coliandKlebsiellaspp. have similar FCA characteristics.

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