Biomaterial Database

An enzyme immunoassay for bovine brucellosis using a monoclonal antibody specific for field strains of Brucella abortus. 1984

M D Gorrell, and G L Milliken, and B J Anderson, and A Pucci

The success of various attempts to discriminate serologically between vaccinated cattle and those infected with field strains of Brucella abortus has been limited. A reliable assay that has this discriminating ability would greatly benefit the Australian Brucellosis Eradication Scheme. Studies of similar diagnostic problems have shown that species and strain-specific monoclonal antibodies with diagnostic potential can be produced. Therefore, the production of monoclonal antibodies specific for Br. abortus field strains was attempted. Spleen cells from Balb/C mice immunized with formalin-killed Br. abortus strain 544 were fused with P3/NSI-1 Ag4-1 cells using polyethylene glycol 4000. The resulting hybrid cells were screened by ELISA using soluble fractions of sonicated, heat-killed (60 degrees C, 1 h) bacteria as antigen. More than 200 cultures of hybrid cells contained detectable amounts of antibody to both field and vaccine strains of Brucella. However, one monoclonal antibody, Br 25, was found to bind field strains of Br. abortus biotypes 1 and 2, but not strain 19, Br. suis, Br. melitensis, or Br. ovis. The titre of Br 25 hybridoma cell culture supernatant is over 2 000, Br 25 cells injected into mice produce ascites fluid having titres of more than 100 000. A sandwich ELISA incorporating successive incubations of Br 25, soluble Brucella antigen, bovine sera and labelled anti-bovine immunoglobulin was developed. Sera from cattle that had been either vaccinated with strain 19 and infected were tested in the ELISA. In this ELISA most infected cattle were positive, whether vaccinated or not, whereas uninfected and vaccinated cattle were negative.

UI	MeSH Term	Description	Entries
D008807	Mice, Inbred BALB C	An inbred strain of mouse that is widely used in IMMUNOLOGY studies and cancer research.	BALB C Mice, Inbred,BALB C Mouse, Inbred,Inbred BALB C Mice,Inbred BALB C Mouse,Mice, BALB C,Mouse, BALB C,Mouse, Inbred BALB C,BALB C Mice,BALB C Mouse
D002002	Brucella	A genus of gram-negative, aerobic bacteria that causes BRUCELLOSIS. Its cells are nonmotile coccobacilli and are animal parasites and pathogens. The bacterium is transmissible to humans through contact with infected dairy products or tissue.
D002003	Brucella abortus	A species of the genus BRUCELLA whose natural hosts are cattle and other bovidae. Abortion and placentitis are frequently produced in the pregnant animal. Other mammals, including humans, may be infected.	Bacterium abortus,Brucella melitensis biovar abortus
D002007	Brucellosis, Bovine	A disease of cattle caused by bacteria of the genus BRUCELLA leading to abortion in late pregnancy. BRUCELLA ABORTUS is the primary infective agent.	Bang's Disease,Bang Disease,Bangs Disease,Bovine Brucelloses,Bovine Brucellosis,Brucelloses, Bovine,Disease, Bang,Disease, Bang's
D002417	Cattle	Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.	Beef Cow,Bos grunniens,Bos indicus,Bos indicus Cattle,Bos taurus,Cow,Cow, Domestic,Dairy Cow,Holstein Cow,Indicine Cattle,Taurine Cattle,Taurus Cattle,Yak,Zebu,Beef Cows,Bos indicus Cattles,Cattle, Bos indicus,Cattle, Indicine,Cattle, Taurine,Cattle, Taurus,Cattles, Bos indicus,Cattles, Indicine,Cattles, Taurine,Cattles, Taurus,Cow, Beef,Cow, Dairy,Cow, Holstein,Cows,Dairy Cows,Domestic Cow,Domestic Cows,Indicine Cattles,Taurine Cattles,Taurus Cattles,Yaks,Zebus
D004797	Enzyme-Linked Immunosorbent Assay	An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed.	ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D006825	Hybridomas	Cells artificially created by fusion of activated lymphocytes with neoplastic cells. The resulting hybrid cells are cloned and produce pure MONOCLONAL ANTIBODIES or T-cell products, identical to those produced by the immunologically competent parent cell.	Hybridoma
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D000907	Antibodies, Bacterial	Immunoglobulins produced in a response to BACTERIAL ANTIGENS.	Bacterial Antibodies
D000911	Antibodies, Monoclonal	Antibodies produced by a single clone of cells.	Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal