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Biomedical subjects

J McNair

Publications and source records attributed to J McNair.

9 recordsLinked to original sources

Development of a sensitive and specific time resolved fluorimetric immunoassay for the bovine acute phase protein haptoglobin (Hp).

Haptoglobin (Hp) is recognised as a major acute phase protein in the bovidae and its presence in serum is used as an indicator of inflammation. A mouse monoclonal antibody (1D9) specific for bovine Hp was labelled with a lanthanide (Eu) chelate and used to develop a competitive immunoassay. This competitive immunoassay allowed direct measurement of Hp in serum and was validated for intra- and interassay coefficients of variation (below 8%). Cross-reactivity with other serum proteins was measured (less than 0.1%) and limits of detection for Hp in serum were established for adult male (0.344 microgram/ml) and adult female cattle (1.589 micrograms/ml). The immunoassay was compared with an established haptoglobin-haemoglobin binding assay.

Animals

Production, preliminary characterisation and applications of monoclonal antibodies to porcine circovirus.

The preparation of monoclonal antibodies (mAbs) to porcine circovirus is described. Preliminary characterisation was carried out on nine mAbs obtained from two fusions and included isotyping, virus neutralisation assays and indirect immunofluorescence staining patterns obtained following immunostaining of both a porcine circovirus (PCV)-persistently infected pig kidney (PK/15/W) and Vero (Vero-PCV) cell line. Significant differences in the staining patterns were observed in both cell lines which appeared to be dependent on the subculture status of the Vero-PCV cultures. The development of a mAb-based antigen capture enzyme linked immunosorbent assay (ELISA) as an aid to virus purification is also described. The use of mAbs for the detection of PCV antigen in cryostat sections from a pig experimentally infected with the virus leading to identification of the sites of replication of PCV is also reported.

Animals

Application of a monoclonal antibody-based enzyme-linked immunosorbent assay for detection of an inflammatory response antigen in subclinical mastitic milk samples.

A monoclonal antibody to a 23.5-kDa bovine inflammatory antigen present in high levels in mastitic milk has been used in an antigen-capture enzyme-linked immunosorbent assay (ELISA) to screen milk samples from herds of cattle for subclinical mastitis. The results from 20 herds with a total of 2,612 quarter samples are presented. Good correlation was observed between the ELISA level and the milk cell count (MCC). The results demonstrated an average of 5% false negatives (1.8% associated with isolates of Staphylococcus aureus and/or Streptococcus spp.) and 7.7% false positives for each herd in relation to mastitic (greater than 400,000 cells per ml) or nonmastitic (less than 400,000 cells per ml) MCCs.

Acetylglucosaminidase

Parent involvement in transition programs.

Parent involvement in programs facilitating the transition of persons with handicaps from school to adult life (work, independent living) was examined. The survey found that a significant number of parents were unaware of any transition program for their offspring, although virtually all parents desired involvement of some kind. Parents also indicated preferences relating to their child not living at home upon graduation from school, working in the community, and earning a wage greater than or equal to the minimum.

Activities of Daily Living

An antigen capture ELISA test using monoclonal antibodies for the detection of Mycoplasma californicum in milk.

The use of monoclonal antibodies to detect Mycoplasma californicum was investigated in an antigen capture microtitre format. The finalized test was highly specific and no cross-reactions were detectable with any of the mastitis associated mycoplasma or bacterial antigens tested. Using a concentration step involving centrifugation, the sensitivity of the test could be improved from 10(5)-10(7) to 10(3)-10(5) colony forming units per ml with pure broth cultures, and from 10(7) to at least 10(6) colony forming units per ml in milk samples from two experimentally infected cows. The antigen detected was partially identified by immunoblotting, which demonstrated two polypeptides of 40 and 46 kD.

Animals

Antigenically important proteins of Aujeszky's disease (pseudorabies) virus identified by immunoblotting.

Immunoblotting was used to identify those Aujeszky's disease virus proteins which elicited major antibody responses in naturally and experimentally infected pigs. Although some proteins comprising purified virus preparations reacted nonspecifically, proteins with mol. wts. of 120 K, 90 K, 71 K and 60 K were antigenically important. These corresponded in size to the virus glycoproteins identified by 3H-glucosamine labelling. Glycoproteins isolated by affinity chromatography from infected PK 15 cells solubilised by Triton X-100 (Triton-soluble glycoproteins) contained antigenic components similar in size to virus glycoproteins and produced minimal non-specific immunoblotting reactions. Antibody responses to the 120 K and 71 K proteins usually occurred together and were more pronounced than responses to other proteins especially at early times postinfection.

Animals

Enzyme-linked immunosorbent assay for detecting antibodies to Aujeszky's disease virus in pigs.

An indirect micro enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to Aujeszky's disease virus in pigs is described. A control antigen prepared from infected cells was included for each serum tested. Of 243 sera from serologically positive farms, 175 (72 per cent) and 147 (60 per cent) were positive by the ELISA test and microtitre serum neutralisation test, respectively. Failure to include a control antigen for each serum would have resulted in 14 sera (6 per cent) being differently recorded. Results for sera from experimental and field infections indicated that seroconversion was more quickly detected by the ELISA test than the microtitre serum neutralisation test. In addition to greater sensitivity the ELISA test has other advantages over the serum neutralisation test. ELISA is a rapid, cheap test which is not dependent on a continuous supply of cell cultures and which can be readily automated.

Animals

Production and preliminary characterization of monoclonal antibodies to chicken anemia agent.

Mice were immunized with partially purified preparations of the Cux-1 isolate of chicken anemia agent (CAA), and their splenocytes were fused with NSO myeloma cells. Three patterns of staining of CAA-infected cells were recognized when the resulting hybridomas were screened by indirect immunofluorescence (IIF). Hybridomas representative of each staining pattern were cloned, and the monoclonal antibodies (MAbs) were characterized. Type 1 staining was indistinguishable from that produced by polyclonal chicken antisera to CAA. Type 2 staining was confined to large nuclear inclusions. Type 3 staining was predominantly nuclear and granular, and differed from type 1 in being more intense and occurring in a higher proportion of nuclei. Three MAbs producing type 1 staining were predominantly Cux-1-specific by IIF; they also reacted to lower titers with the Gifu-1 isolate but not at all with three other CAA isolates. These MAbs had very slight neutralizing activity against Cux-1. Another MAb giving type 1 staining reacted with all CAA isolates tested to high titers in IIF and neutralization tests. MAbs with type 2 and type 3 staining reacted by IIF with all CAA isolates tested but possessed no neutralizing activity. The availability of MABs to CAA should facilitate development of diagnostic tests for the virus.

Animals