scholarly journals EXPRESSION OF RECOMBINANT P24 PROTEIN IN ESCHERICHIA COLI FOR SEROLOGICAL DIAGNOSIS OF BOVINE LEUKOSIS

2021 ◽  
pp. 41-48
Author(s):  
Baltin K ◽  
Shustov A ◽  
Khassenov B.
Keyword(s):  
Escherichia Coli ◽  
Immunosorbent Assay ◽  
Leukemia Virus ◽  
Total Yield ◽  
Plasmid Vector ◽  
Positive Control ◽  
Lymphoproliferative Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Tests ◽  
P24 Protein

Bovine leukemia is an infectious lymphoproliferative disease in cattle caused by the bovine leukemia virus (BLV). Production losses associated with BLV include increased cull rates, decreased milk production, or reduced fertility. Serological tests are used for diagnosis of BLV, of which the most sensitive is enzyme-linked immunosorbent assay (ELISA). Capsid p24 protein is a diagnostically important antigen of the BLV. The gene of p24 protein was cloned into pET-28 plasmid vector. The recombinant p24 antigen was obtained by using Escherichia coli ArcticExpressRP(DE3)_pET-28/р24 strain. The total yield of the purified p24 protein was 76 mg. Testing of the recombinant p24 protein on control sera showed that the p24 binds to positive control sera and has high antigenic activity. The pilot testing of the recombinant p24 protein on 48 animal samples resulted in 83% positive and 17% negative. The results indicate that the recombinant p24 protein is promising in enzyme-linked immunosorbent assay for the diagnosis of bovine leukemia.

scholarly journals Detection of lymphoproliferative disease virus by an enzymelinked immunosorbent assay

1987 ◽  
Vol 99 (3) ◽  
pp. 711-722 ◽  
Author(s):  
J. R. Patel ◽  
R. W. Shilleto
Keyword(s):  
Disease Virus ◽  
Immunosorbent Assay ◽  
High Speed ◽  
Reverse Transcriptase Activity ◽  
Lymphoproliferative Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Tests ◽  
Enzymelinked Immunosorbent Assay ◽  
Specific Antigens ◽  
Avian Sarcoma

SUMMARYHitherto, detection of lymphoproliferative disease virus (LPDV), a C-type retrovirus of turkeys, has proved difficult since no tissue culture or serological assay has been available. Development of serological tests has been hampered by the problems of raising virus-specific antisera. An indirect enzyme-linked immunosorbent assay (ELISA) is reported, using a viral antiserum raised with bromelain-digested virus.The assay specifically detected purified virus at a concentration of 250 ng/ml or greater. In an experiment to detect virus in plasma from turkeys over a period of 4 weeks following LPDV infection, ELISA results correlated closely with the viral reverse transcriptase activity. Both assays were of similar sensitivity and detected small amounts of virus in high-speed pellets of turkey plasma. Evidence is presented indicating that LPDV-infected or hyperimmunized turkeys do not produce readily detectable circulating viral antibodies. In reciprocal ELISA tests, using antibodies to group-specific antigens of other avian retrovirus groups (avian sarcoma-leukosis (ASLV) and reticuloendotheliosis (REV)) no antigenic cross-reaction was found between LPDV, ASLV and REV.

scholarly journals Unreliable Measurement of Basophil Maximum Leukotriene Release with the Bühlmann CAST 2000 Enzyme-Linked Immunosorbent Assay Kit

2006 ◽  
Vol 13 (3) ◽  
pp. 420-422 ◽  
Author(s):  
S. E. Burastero ◽  
C. Paolucci ◽  
D. Breda ◽  
G. Monasterolo ◽  
R. E. Rossi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
False Positive ◽  
Positive Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Positive Results

ABSTRACT The Bühlmann CAST 2000 enzyme-linked immunosorbent assay is a potentially useful assay for measuring sulfidoleukotrienes released in vitro by allergen-challenged basophils. However, we observed that the positive-control reagent yielded positive signals in cell-free systems. These false-positive results depended on using a mouse anti-FcεRI monoclonal antibody and were prevented by degranulation-inducing reagents other than mouse monoclonal antibodies.

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.

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