scholarly journals Multiplex Screening of Surface Proteins from Mycoplasma mycoides subsp. mycoides Small Colony for an Antigen Cocktail Enzyme-Linked Immunosorbent Assay

2009 ◽  
Vol 16 (11) ◽  
pp. 1665-1674 ◽  
Author(s):  
Maja Neiman ◽  
Carl Hamsten ◽  
Jochen M. Schwenk ◽  
Göran Bölske ◽  
Anja Persson
Keyword(s):  
Immunosorbent Assay ◽  
False Negative ◽  
Negative Control ◽  
Surface Proteins ◽  
Enzyme Linked Immunosorbent Assay ◽  
Contagious Bovine Pleuropneumonia ◽  
Mycoplasma Mycoides ◽  
Small Colony ◽  
Suspension Bead Array ◽  
Selection Of

ABSTRACT A recombinant antigen cocktail enzyme-linked immunosorbent assay (ELISA) for diagnosis of contagious bovine pleuropneumonia (CBPP) was developed after careful selection of antigens among one-third of the surface proteome proteins of the infectious agent Mycoplasma mycoides subsp. mycoides small colony (M. mycoides SC). First, a miniaturized and parallelized assay system employing antigen suspension bead array technology was used to screen 97 bovine sera for humoral immune responses toward 61 recombinant surface proteins from M. mycoides SC. Statistical analysis of the data resulted in selection of eight proteins that showed strong serologic responses in CBPP-affected sera and minimal reactivity in negative control sera, with P values of <10−6. Only minor cross-reactivity to hyperimmune sera against other mycoplasmas was observed. When applied in an ELISA, the cocktail of eight recombinant antigens allowed a fivefold signal separation between 24 CBPP-affected and 23 CBPP-free sera from different geographical origins. No false-positive results and only two false-negative results were obtained. In conclusion, the selected recombinant mycoplasma antigens qualified as highly specific markers for CBPP and could be employed in both a suspension bead array platform and a cocktail ELISA setting. This set of proteins and technologies therefore offers a powerful combination to drive and further improve serological assays toward reliable, simple, and cost-effective diagnosis of CBPP.

scholarly journals Development of a Novel Cocktail Enzyme-Linked Immunosorbent Assay and a Field-Applicable Lateral-Flow Rapid Test for Diagnosis of Contagious Bovine Pleuropneumonia

2016 ◽  
Vol 54 (6) ◽  
pp. 1557-1565 ◽  
Author(s):  
Martin Heller ◽  
Nimmo Gicheru ◽  
Georgina Tjipura-Zaire ◽  
Cecilia Muriuki ◽  
Mingyan Yu ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Rapid Test ◽  
Lateral Flow ◽  
Sub Saharan Africa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Contagious Bovine Pleuropneumonia ◽  
Serum Samples ◽  
Content Type ◽  
Mycoplasma Mycoides ◽  
Sub Saharan

Contagious bovine pleuropneumonia (CBPP) is a severe respiratory disease that is widespread in sub-Saharan Africa. It is caused byMycoplasma mycoidessubsp.mycoides, a bacterium belonging to theMycoplasma mycoidescluster. In the absence of an efficient CBPP vaccine, improved and easy-to-use diagnostic assays for recurrent testing combined with isolation and treatment of positive animals represent an option for CBPP control in Africa. Here we describe the comprehensive screening of 17 immunogenicMycoplasma mycoidessubsp.mycoidesproteins using well-characterized bovine sera for the development of a novel cocktail enzyme-linked immunosorbent assay (ELISA) for laboratory use. Two recombinantMycoplasmaimmunogens, MSC_0136 and MSC_0636, were used to set up a standardized cocktail ELISA protocol. According to the results from more than 100 serum samples tested, the sensitivity and specificity of the novel cocktail ELISA were 85.6% and 96.4%, respectively, with an overall diagnostic accuracy comparable to that of the Office International des Epizooties (OIE)-prescribed serological assays. In addition, we provide a proof of principle for a field-applicable, easy-to-use commercially produced prototype lateral-flow test for rapid (<30-min) diagnosis of CBPP.

scholarly journals Rapid Detection of Campylobacter coli, C. jejuni, and Salmonella enterica on Poultry Carcasses by Using PCR-Enzyme-Linked Immunosorbent Assay

2003 ◽  
Vol 69 (6) ◽  
pp. 3492-3499 ◽  
Author(s):  
Yang Hong ◽  
Mark E. Berrang ◽  
Tongrui Liu ◽  
Charles L. Hofacre ◽  
Susan Sanchez ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Salmonella Enterica ◽  
Rapid Detection ◽  
Diarrheal Disease ◽  
False Negative ◽  
Enzyme Linked Immunosorbent Assay ◽  
Campylobacter Coli ◽  
Culture Methods ◽  
Isolation And Identification ◽  
Serological Tests

ABSTRACT Contamination of retail poultry by Campylobacter spp. and Salmonella enterica is a significant source of human diarrheal disease. Isolation and identification of these microorganisms require a series of biochemical and serological tests. In this study, Campylobacter ceuE and Salmonella invA genes were used to design probes in PCR-enzyme-linked immunosorbent assay (ELISA), as an alternative to conventional bacteriological methodology, for the rapid detection of Campylobacter jejuni, Campylobacter coli, and S. enterica from poultry samples. With PCR-ELISA (40 cycles), the detection limits for Salmonella and Campylobacter were 2 � 102 and 4 � 101 CFU/ml, respectively. ELISA increased the sensitivity of the conventional PCR method by 100- to 1,000-fold. DNA was extracted from carcass rinses and tetrathionate enrichments and used in PCR-ELISA for the detection of Campylobacter and S. enterica, respectively. With PCR-ELISA, Salmonella was detected in 20 of 120 (17%) chicken carcass rinses examined, without the inclusion of an enrichment step. Significant correlation was observed between PCR-ELISA and cultural methods (kappa = 0.83; chi-square test, P < 0.001) with only one false negative (1.67%) and four false positives (6.67%) when PCR-ELISA was used to screen 60 tetrathionate enrichment cultures for Salmonella. With PCR-ELISA, we observed a positive correlation between the ELISA absorbance (optical density at 405 nm) and the campylobacter cell number in carcass rinse, as determined by standard culture methods. Overall, PCR-ELISA is a rapid and cost-effective approach for the detection and enumeration of Salmonella and Campylobacter bacteria on poultry.

A simplified PCR method for genotyping Mycoplasma mycoides subspecies mycoides small colony: The aetiologic agent of contagious bovine pleuropneumonia

2012 ◽  
Vol 159 (1-2) ◽  
pp. 257-259
Author(s):  
Colin P. Churchward ◽  
Miroslav Hlúšek ◽  
Robin A.J. Nicholas ◽  
Roger D. Ayling ◽  
Laura McAuliffe
Keyword(s):  
Contagious Bovine Pleuropneumonia ◽  
Mycoplasma Mycoides ◽  
Small Colony ◽  
Pcr Method

scholarly journals Selection of single chain antibody fragments binding to the extracellular domain of 4-1BB receptor by phage display technology

Tumor Biology ◽  
2017 ◽  
Vol 39 (3) ◽  
pp. 101042831769592 ◽  
Author(s):  
Salman Bagheri ◽  
Mehdi Yousefi ◽  
Elmira Safaie Qamsari ◽  
Farhad Riazi-Rad ◽  
Mohsen Abolhassani ◽  
...  
Keyword(s):  
Phage Display ◽  
Immunosorbent Assay ◽  
Specific Binding ◽  
Interleukin 2 ◽  
Extracellular Domain ◽  
Surface Glycoprotein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Single Chain ◽  
Single Chain Variable Fragments ◽  
Selection Of

The 4-1BB is a surface glycoprotein that pertains to the tumor necrosis factor–receptor family. There is compelling evidence suggesting important roles for 4-1BB in the immune response, including cell activation and proliferation and also cytokine induction. Because of encouraging results of different agonistic monoclonal antibodies against 4-1BB in the treatment of cancer, infectious, and autoimmune diseases, 4-1BB has been suggested as an attractive target for immunotherapy. In this study, single chain variable fragment phage display libraries, Tomlinson I+J, were screened against specific synthetic oligopeptides (peptides I and II) designed from 4-1BB extracellular domain. Five rounds of panning led to selection of four 4-1BB specific single chain variable fragments (PI.12, PI.42, PII.16, and PII.29) which showed specific reaction to relevant peptides in phage enzyme-linked immunosorbent assay. The selected clones were successfully expressed in Escherichia coli Rosetta-gami 2, and their expression was confirmed by western blot analysis. Enzyme-linked immunosorbent assay experiments indicated that these antibodies were able to specifically recognize 4-1BB without any cross-reactivity with other antigens. Flow cytometry analysis demonstrated an acceptable specific binding of the single chain variable fragments to 4-1BB expressed on CCRF-CEM cells, while no binding was observed with an irrelevant antibody. Anti-4-1BB single chain variable fragments enhanced surface CD69 expression and interleukin-2 production in stimulated CCRF-CEM cells which confirmed the agonistic effect of the selected single chain variable fragments. The data from this study have provided a rationale for further experiments involving the biological functions of anti-4-1BB single chain variable fragments in future studies.

High Prevalence of False-Negative Anti-HTLV Type I/II Enzyme-Linked Immunosorbent Assay Results in HIV Type 1-Positive Patients

1997 ◽  
Vol 13 (13) ◽  
pp. 1141-1146 ◽  
Author(s):  
GIANGUGLIELMO ZEHENDER ◽  
CHIARA DE MADDALENA ◽  
MONICA GIANOTTO ◽  
BARBARA CAVALLI ◽  
SARA SANTAMBROGIO ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
False Negative ◽  
Enzyme Linked Immunosorbent Assay ◽  
Type I ◽  
High Prevalence ◽  
Hiv Type 1

scholarly journals An enzyme-linked immunosorbent assay for platelet compatibility testing

Blood ◽  
1983 ◽  
Vol 62 (4) ◽  
pp. 744-749 ◽  
Author(s):  
JD Tamerius ◽  
JG Curd ◽  
P Tani ◽  
R McMillan
Keyword(s):  
Antibody Titer ◽  
Immunosorbent Assay ◽  
Clinical Laboratory ◽  
Clinical Problem ◽  
Enzyme Linked Immunosorbent Assay ◽  
Microtiter Plates ◽  
Volunteer Donor ◽  
Antibody Titers ◽  
Test Serum ◽  
Selection Of

Abstract The selection of platelet donors for patients who are refractory to random donor platelets often presents a difficult clinical problem. We describe an enzyme-linked immunosorbent assay (ELISA) for evaluating alloantibodies in refractory patients. Platelets from prospective donors are immobilized on microtiter plates and, after incubation with test serum and washing, platelet-bound IgG is detected with enzyme- linked anti-human IgG. Platelets from 46 prospective donors were tested. Twenty-two were judged compatible (reciprocal of the antibody titer less than 16) and, of these, 15 were used as platelet donors; each gave a measurable platelet increment after transfusion. The magnitude of the response was roughly proportional to the assay results. Platelets from donors giving antibody titers less than 4 resulted in platelet increments at 1 hr ranging from 4,890 to 22,200 (median 12,600), while platelets from donors giving titers of 8 or 16 resulted in lesser increments (550–4548). Conversely, 5 of the 24 patients found incompatible by the assay (titer greater than 16) gave no platelet increment, and in 3 instances, the recipient developed fever and chills after the transfusion. The assay is sensitive, simple, and adaptable to the clinical laboratory. Platelets from volunteer donor panels can be plated and stored for up to 6 mo.

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