Detection of Group D Salmonellae Including Salmonella Enteritidis in Eggs by Polymyxin-Based Enzyme-Linked Immunosorbent Assay

2008 ◽  
Vol 71 (2) ◽  
pp. 392-396 ◽  
Author(s):  
BURTON W. BLAIS ◽  
AMALIA MARTINEZ-PEREZ
Keyword(s):  
Immunosorbent Assay ◽  
Salmonella Enteritidis ◽  
Effective Means ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Selective Enrichment ◽  
Affinity Adsorbent ◽  
Egg Products ◽  
Soya Peptone ◽  
Group D

A high-throughput, rapid method was devised for the detection of Salmonella Enteritidis in egg products. For each target organism, preenrichment in nutrient broth was followed by selective enrichment in Rappaport-Vassiliadis soya peptone and tetrathionate brilliant green broths or by plating on modified semisolid Rappaport Vassiliadis (MSRV) agar medium. The presence of Salmonella Enteritidis was determined by subjecting portions of the selective broth cultures or swarming growth on MSRV medium to an enzyme-linked immunosorbent assay (ELISA) procedure using polymyxin immobilized in the wells of a microtiter plate as a high-affinity adsorbent for lipopolysaccharide (LPS) antigens. Sample extracts were reacted with polymyxin-coated microwells, and captured LPS antigens were detected immunoenzymatically with a commercially available Salmonella factor O9–specific antibody. The polymyxin-ELISA was 100% sensitive and 100% specific for Salmonella strains bearing the O9 antigen. When the ELISA was combined with enrichment using either the selective broths or plating on MSRV medium, the system was an effective means for detection of Salmonella Enteritidis in artificially inoculated egg products. The polymyxin-ELISA is a simple and inexpensive assay for group D salmonellae (including Salmonella Enteritidis) in a convenient 96-well microtiter plate format, making this system ideally suited for processing large numbers of samples.

Detection of Escherichia coli O157 in Foods by a Novel Polymyxin-Based Enzyme-Linked Immunosorbent Assay

2005 ◽  
Vol 68 (2) ◽  
pp. 233-238 ◽  
Author(s):  
BURTON W. BLAIS ◽  
JOHANNA LEGGATE ◽  
JESSICA BOSLEY ◽  
AMALIA MARTINEZ-PEREZ
Keyword(s):  
Escherichia Coli ◽  
Immunosorbent Assay ◽  
Microtiter Plate ◽  
Complete Agreement ◽  
Enzyme Linked Immunosorbent Assay ◽  
Escherichia Coli O157 ◽  
Elisa System ◽  
Culture Techniques ◽  
E Coli ◽  
Affinity Adsorbent

An enzyme-linked immunosorbent assay (ELISA) system was developed using polymyxin immobilized in the wells of a microtiter plate as a high-affinity adsorbent for Escherichia coli O157 lipopolysaccharide (LPS) antigens. Extracts from cell suspensions were reacted with polymyxin-coated microwells followed by immunoenzymatic detection of captured LPS antigens using a commercially available anti–E. coli O157 antibody–peroxidase conjugate and a 3,3′,5,5′-tetramethylbenzidine substrate. The polymyxin ELISA was highly sensitive and specific for E. coli strains bearing the O157 antigen. When this ELISA was combined with enrichment, results were in complete agreement with those of standard culture techniques for the detection of this pathogen in a variety of artificially inoculated and naturally contaminated foods. The polymyxin ELISA is a simple and inexpensive assay for E. coli O157 with a 96-well microtiter plate format, making this system ideally suited for processing large numbers of samples.

scholarly journals Enzyme-linked immunosorbent assay with a Salmonella enteritidis antigen for differentiating infected from vaccinated poultry

2000 ◽  
Vol 31 (5) ◽  
pp. 491-497 ◽  
Author(s):  
Cristina Solano ◽  
Julia Galindo ◽  
Bego�a Sesma ◽  
Miguel Alvarez ◽  
Mar�a J. Solsona ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Salmonella Enteritidis ◽  
Enzyme Linked Immunosorbent Assay

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.

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