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At least 595 records · Page 33Linked to original sources
PubMed1999

[Multiplex PCR for the diagnostic detection of Coxiella burnetii in cow's milk].

A multiplex PCR based assay was developed for the highly sensitive and specific detection of Coxiella (C.) burnetii in cow's milk. The assay simultaneously amplifies a diagnostic target within the C. burnetii IS1111 sequence and a control target within the bovine CD18 gene. The internal PCR amplification control allows the discrimination of false negative results (single tube reaction failures) from negative results due to true absence of target sequences. In order to maximize the sensitivity of the assay, a sample preparation method including a centrifugation step to concentrate the bacterium was developed. In milk samples artificially contaminated with serial dilutions of C. burnetii, about four particles per ml could reproducibly be detected. The sensitivities of both assays, multiplex PCR and PCR with only a single pair of primers ('simplex' PCR), were observed to be similar.

Animals↗
PubMed1996

Improved method of isolation of Coxiella burnetii proteins.

An improved method of isolation of Coxiella burnetti proteins was developed. It consists of a combination of detergent (sodium dodecyl sulphate (SDS) or sodium deoxycholate (DOC) and hot phenol treatments. The resulting phenol phase (PP) contained either lipopolysaccharide-(LPS) free proteins (DOC extraction) or proteins contaminated with LPS (SDS extraction), while the water phase (WP) contained LPS. Isolated C. burnetii proteins induced in mice and rabbits antibodies reacting in immunoblot analysis with both phase I and II C. burnetii corpuscles. A rabbit serum against C. burnetii prepared by DOC-phenol extraction did not react with purified I C. burnetii LPS in immunoblot analysis.

Animals↗
PubMed1987

Detection of Coxiella burnetii by the immunoperoxidase technique.

An immunoperoxidase method using Coxiella (C.) burnetii-specific hyperimmune serum raised in rabbits, and swine anti-rabbit horseradish peroxidase conjugate was applied to visualize C. burnetii in BGM cell cultures. The technique proved to be highly specific and did not result in any unspecific background staining. The technique was applied successfully to study the organism during multiplication in vacuoles of epithelial cells and in the phagolysosomes of macrophages. Preliminary results further indicate that the technique may be useful for the detection of the pathogen in tissues and milk samples.

Animals↗
PubMed1996

Structural study on a lipopolysaccharide from Coxiella burnetii strain Nine Mile in avirulent phase II.

A lipopolysaccharide isolated from Coxiella burnetti strain Nine Mile in avirulent phase II contains in the lipid A proximal region D-mannose, D-glycero-D-manno-heptose, and 3-deoxy-D- manno-oct-2-ulosonic acid (Kdo) in the molar ratio 2:2:3. The primary structure 1 of the heptasaccharide was determined by glycose analysis, methylation analysis, ESI-MS, and FABMS. [sequence: see text]

Carbohydrate Conformation↗
PubMed1989

[Immunogenic and immunomodulating properties of phospholipids from Coxiella burnetii].

It is proved that the challenging of the extract isolated from Coxiella (as a result of treatment with the mixture of chloroform and methanol) to animals doesn't provoke the appearance of antibodies to these microorganisms. It also doesn't cause the sensibilization of lymphocytes to Coxiella, but the extract can interact specifically with a serum containing antibodies to Coxiella. It permits to characterise the tested extract of Coxiella as a hapten with properties of adjuvant because it stimulates antibody-appearance.

Adjuvants, Immunologic↗
PubMed1993

Plasmid based differentiation and detection of Coxiella burnetii in clinical samples.

A "nested" PCR approach with primers based on conserved plasmid sequences was used for the highly sensitive and specific detection of Coxiella (C.) burnetii in clinical samples collected from cattle, dogs, cats and humans. Results were in good agreement with those obtained from Capture-ELISA and isolation of the organism in BGM cell culture. We also tested primers with sequences derived from genomic DNA and sequences based on 16S rRNA. In addition, we applied PCR for the differentiation of C. burnetii plasmid types from 28 isolates originating from the USA, Europe and South Africa. Reference isolates Nine Mile RSA493, Dugway 5J108-111 and all European isolates tested were recognized only by primers specific for the QpH1 plasmid. One isolate from a goat abortion in Namibia reacted identically to the reference isolate Priscilla Q177 bearing the QpRS plasmid. Reference isolate S Q217 with plasmid sequences integrated into the genome reacted with none of the plasmid-specific primer pairs.

Animals↗
PubMed1992

Monoclonal antibody based capture ELISA/ELIFA for detection of Coxiella burnetii in clinical specimens.

A CAPTURE ELISA/ELIFA system based on monoclonal capture and biotinylated monoclonal detection antibody is described. The assay is fast, highly specific and detects a minimum dose of 2500 Coxiella (C.) burnetii particles. In contrast to the sophisticated and cumbersome isolation procedures, even non-specialized laboratories could use this assay system for investigating clinical samples of different origin for C. burnetii within a short period of time.

Antibodies, Monoclonal↗