Participation of peptide moieties in adhesive behavior of antigenic mannans of Candida albicans to the plastic microtiter plate in enzyme-linked immunosorbent assay.

1987 ◽  
Vol 33 (10) ◽  
pp. 1925-1928 ◽  
Author(s):  
M Tojo ◽  
N Shibata ◽  
T Mikami ◽  
M Suzuki ◽  
S Suzuki
Keyword(s):  
Candida Albicans ◽  
Immunosorbent Assay ◽  
Alkaline Solution ◽  
Yeast Species ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Aqueous Alkaline Solution ◽  
Pathogenic Yeast ◽  
Color Development ◽  
Polystyrene Microtiter Plate

Abstract We report our studies of the mechanism of the adhesion of mannans of Candida albicans NIH A-207 and C. albicans NIH B-792 strains to the surface of a polystyrene microtiter plate being utilized for enzyme-linked immunosorbent assay (EIA) of antigens of this pathogenic yeast species. This binding was manifested predominantly by the peptide moieties of the mannans forming hydrophobic bonds with the plastic molecule. Eliminating the peptide of each mannan by treating it with a hot aqueous alkaline solution of NaBH4 also eliminated color development in the EIA.

Prostatic inhibin-like peptide quantified in urine of prostatic cancer patients by enzyme-linked immunosorbent assay.

1989 ◽  
Vol 35 (7) ◽  
pp. 1376-1379 ◽  
Author(s):  
T R Teni ◽  
A H Bandivdekar ◽  
A R Sheth ◽  
N A Sheth
Keyword(s):  
Cancer Patients ◽  
Immunosorbent Assay ◽  
Prostatic Cancer ◽  
Solid Phase ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Recovery ◽  
The Status ◽  
Polystyrene Microtiter Plate ◽  
The Mean

Abstract This is a highly specific enzyme-linked immunosorbent assay (ELISA) for measuring prostatic inhibin-like peptide (PIP) in urine, in which we use penicillinase (EC 3.5.2.6) conjugated with PIP and, as solid phase, a polystyrene microtiter plate. We used this ELISA to measure PIP in 24-h urine specimens from men with prostatic cancer (PCa) and from age-matched controls. For prostatic cancer patients the mean +/- SEM urinary PIP of 36.1 +/- 5 micrograms/24 h was significantly (P less than 0.001) lower than the mean of 127.1 +/- 9 micrograms/24 h for the age-matched controls. PIP values for 30 samples measured by both ELISA and RIA correlated well (r = 0.985). We could detect as little as 1.56 ng of PIP in a sample. Analytical recovery of added PIP ranged from 91% to 104%. Mean CVs were 8.9% within-assay and 12.7% between-assay. We believe that this ELISA will be useful in assessing the status of PIP in men with normal and diseased prostates and in examining the function of the hypothalamus-pituitary-prostate axis.

Competitive Direct Enzyme-Linked Immunosorbent Assay for Detection of Sulfamethazine Residues in Swine Urine and Muscle Tissue

1988 ◽  
Vol 71 (6) ◽  
pp. 1137-1140 ◽  
Author(s):  
Deborah E Dixon-Holland ◽  
Stanley E Katz
Keyword(s):  
Horseradish Peroxidase ◽  
Muscle Tissue ◽  
Immunosorbent Assay ◽  
Microtiter Plate ◽  
Test System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sensitive Assay ◽  
Saline Extract ◽  
Swine Urine ◽  
Phosphate Buffered Saline

Abstract A sensitive assay for the detection of sulfamethazine in swine urine and muscle tissue using a direct competitive enzyme-linked immunosorbent assay (ELISA) has been developed. Undiluted urine or a phosphate-buffered saline extract of pork muscle tissue is mixed with an enzyme-labeled conjugate of sulfamethazine and horseradish peroxidase. The mixture is added to wells of a microtiter plate coated with antibody to sulfamethazine. After the test system is incubated, washed, and re-incubated with substrate and the reaction is stopped, the absorbance is measured at 405 nm. Levels of sulfamethazine as low as 20 ng sulfamethazine/g muscle tissue and 10 ng sulfamethazine/ mL swine urine were detected and estimated

Highly Sensitive, Specific Enzyme-Linked Immunosorbent Assay of Neopterin and Biopterin in Biological Samples

1992 ◽  
Vol 38 (10) ◽  
pp. 1954-1958 ◽  
Author(s):  
S Ogiwara ◽  
K Kiuchi ◽  
T Nagatsu ◽  
R Teradaira ◽  
I Nagatsu ◽  
...  
Keyword(s):  
Human Serum ◽  
Immunosorbent Assay ◽  
Lupus Erythematosus ◽  
Present Method ◽  
High Performance ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Usual Manner ◽  
Healthy Control ◽  
Antigen Antibody

Abstract An enzyme immunosorbent assay of neopterin and biopterin on a polystyrene microtiter plate has been developed. A conjugate of neopterin or biopterin to bovine serum albumin was used to raise a specific antiserum against neopterin or biopterin in rabbits. An incubation mixture of the antiserum and samples prepared from human serum underwent another antigen-antibody reaction with the hapten fixed on the microtiter plate. The amount of antibody bound to the fixed hapten, which is inverse to the amount of hapten in the sample, was determined by using anti-rabbit IgG-horseradish peroxidase conjugate in a usual manner by measuring absorbance at 490 nm after reaction with o-phenylenediamine and hydrogen peroxide. The minimal detectable amounts of neopterin and biopterin were approximately 0.1 pmol. The specificity of the assay was so high that the assay system for neopterin completely distinguished it from biopterin, as judged from the cross-reaction of 0.002%, and vice versa. The amounts of neopterin and biopterin in human serum determined by the present method agreed well with those determined by high-performance liquid chromatography. We used the present method to determine the concentrations of neopterin in serum from healthy control subjects and patients with cancers and systemic lupus erythematosus; the results were consistent with literature data.

A High Throughput Enzyme-Linked Immunosorbent Assay for Inhibitors of the Interaction Between Retinoblastoma Protein and the Leu-X-Cys-X-Glu Motif

1997 ◽  
Vol 2 (4) ◽  
pp. 207-211 ◽  
Author(s):  
Victoria Alice Ellsmore ◽  
Al Peng Teoh ◽  
Arasu Ganesan
Keyword(s):  
Immunosorbent Assay ◽  
High Throughput ◽  
High Throughput Screening ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Retinoblastoma Tumor Suppressor ◽  
Multiple Antigenic Peptide ◽  
Peptide Map ◽  
Retinoblastoma Tumor ◽  
Lxcxe Motif

A 96-well enzyme-linked immunosorbent assay was developed to discover compounds that inhibit the binding of the Leu-X-Cys-X-Glu (LXCXE) motif to the retinoblastoma tumor suppressor protein (pRB). The assay uses a LXCXE-containing multiple antigenic peptide (MAP) which is immobilized on a microtiter plate. A truncated form of pRB is added and the amount bound detected by a monoclonal antibody. This rapid assay was employed in high throughput screening of crude natural product extracts and discrete compounds.

scholarly journals Evaluation of an Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assay for Diagnosis of Orientia tsutsugamushi Infection

2003 ◽  
Vol 10 (3) ◽  
pp. 394-398 ◽  
Author(s):  
Won-Jong Jang ◽  
Myung-Suk Huh ◽  
Kyung-Hee Park ◽  
Myung-Sik Choi ◽  
Ik-Sang Kim
Keyword(s):  
Immunosorbent Assay ◽  
Scrub Typhus ◽  
Microtiter Plate ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Orientia Tsutsugamushi ◽  
Serum Samples ◽  
Capture Elisa ◽  
Igm Antibodies ◽  
Immunodominant Antigen

ABSTRACT To differentiate scrub typhus from other acute febrile diseases, a rapid and reliable serological diagnosis is important. We developed an immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) for diagnosis of recent Orientia tsutsugamushi infections in humans. The 56-kDa major outer membrane protein of O. tsutsugamushi is well known as the most immunodominant antigen in scrub typhus. The test is based on the use of the biotinylated recombinant 56-kDa protein of O. tsutsugamushi Boryong, Bor56, which was expressed as a fusion protein with a maltose-binding protein in Escherichia coli. In the test, the serum IgM antibodies were captured by anti-human IgM antibodies coated onto a microtiter plate. The captured IgM antibodies were revealed through sequential addition of biotinylated Bor56 antigen and peroxidase-conjugated streptavidin to the plate. The IgM capture ELISA was compared with the immunofluorescence antibody assay (IFA) by testing 176 serum samples from patients with diagnosed cases of rickettsial disease and patients with other acute febrile diseases. Of the 81 IgG IFA-positive samples, 78 tested positive (sensitivity, 96.3%) and all 31 IgM IFA-positive samples tested positive (sensitivity, 100%) by the IgM capture ELISA. The specificity of the IgM capture ELISA was 99%, and 1 of the 95 IFA-negative samples was positive in the assay. These results strongly suggest that IgM capture ELISA using the recombinant Bor56 antigen is a reliable and detailed method for the detection of early O. tsutsugamushi infection.

scholarly journals The Hyphal and Yeast Forms of Candida albicans Bind the Complement Regulator C4b-Binding Protein

2004 ◽  
Vol 72 (11) ◽  
pp. 6633-6641 ◽  
Author(s):  
T. Meri ◽  
A. M. Blom ◽  
A. Hartmann ◽  
D. Lenk ◽  
S. Meri ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Confocal Microscopy ◽  
Binding Site ◽  
Binding Protein ◽  
Alternative Pathway ◽  
Factor H ◽  
Enzyme Linked Immunosorbent Assay ◽  
Classical Pathway ◽  
Pathogenic Yeast ◽  
Yeast Surface

ABSTRACT Candida albicans, an important pathogenic yeast, activates all three pathways of the complement system. To understand how this yeast evades the effects of the activated system, we have analyzed the binding of the classical pathway inhibitor C4b-binding protein (C4BP) by C. albicans. Purified native as well as recombinant C4BP bound dose dependently to the yeast and hyphal forms, as shown by multiple methods, such as confocal microscopy, flow cytometry, a novel enzyme-linked immunosorbent assay, absorption from human serum, and direct binding assays with purified proteins. A prominent binding site was identified at the tip of the germ tube, a structure that is considered important for tissue penetration and pathogenesis. The binding site in C4BP was localized to the two N-terminal complement control protein domains by using recombinant deletion constructs and site-specific monoclonal antibodies. As the alternative pathway inhibitors factor H and FHL-1 also bind to C. albicans, the binding of all three plasma proteins was compared. Simultaneous binding of the classical regulator C4BP and the alternative pathway regulator factor H was demonstrated by confocal microscopy. In addition, FHL-1 competed for binding with C4BP, suggesting that these two related complement regulators bind to the same structures on the yeast surface. The surface-attached C4BP maintains its complement regulatory activities and inactivates C4b. The surface-attached human C4BP serves multiple functions relevant for immune evasion and likely pathogenicity. It inhibits complement activation at the yeast surface and, in addition, mediates adhesion of C. albicans to host endothelial cells.

scholarly journals Evaluation of a Novel Enzyme-Linked Immunosorbent Assay To Detect Immunoglobulin G Antibody to Enolase for Serodiagnosis of Invasive Candidiasis

2007 ◽  
Vol 14 (3) ◽  
pp. 318-319 ◽  
Author(s):  
Ana Laín ◽  
María D. Moragues ◽  
Juan Carlos García Ruiz ◽  
Joaquín Mendoza ◽  
Ana Camacho ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Immunosorbent Assay ◽  
Immunoglobulin G ◽  
Invasive Candidiasis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Predictive Values ◽  
Sensitivity Specificity ◽  
Immunocompetent Patients

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.

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