Enzyme-linked immunosorbent fluorescence assay and high-pressure liquid chromatography for analysis of humoral immune responses to Coxiella burnetti proteins

Section: Discussionmentioning

confidence: 99%

Characterization of the Coxiella burnetii sucB Gene Encoding an Immunogenic Dihydrolipoamide Succinyltransferase

Nguyen

Yamaguchi

et al. 1999

“…14, 23, 25). The lipopolysaccharides (25) and surface proteins (15,17) were also used as target antigens. These antigens are usually derived from purified organisms.…”

mentioning

confidence: 99%

“…Previous studies indicated that sera taken from vaccinated cattle and acutely infected guinea pigs included antibodies to a 27-kDa outer membrane protein (OMP) (3,4,15,17). The 27-kDa OMP is suggested to be related to the pathogenesis and protective immunity of host animals (15,17).…”

mentioning

confidence: 99%

Evaluation of a Recombinant 27‐kDa Outer Membrane Protein of Coxiella burnetii as an Immunodiagnostic Reagent

Zhang

Hotta

et al. 1998

The 27-kDa outer membrane protein from eight strains of Coxiella burnetii was expressed in the pET-21c protein expression system. Two fusion proteins with molecular masses of 30 and 32 kDa were evident in all eight of the recombinants by SDS-PAGE and immunoblotting. A protein having an approximate size of 30 kDa was purified from the Escherichia coli lysates by one-step affinity purification. The utility of the purified recombinant protein in ELISA was also evaluated by testing its reactivity with human sera and comparing this reactivity with that of Nine Mile phase II antigen. All of the 40 IF-positive serum samples were ELISA-positive for both the Nine Mile phase II and recombinant antigens, and negative serum controls were negative for both antigens. These results suggest that ELISA with the 27-kDa recombinant antigen is a sensitive and specific method for detecting anti-C. burnetii antibodies in human sera.

“…A 27-kDa protein has been recognized in guinea pigs early in the course of infection and following vaccination (16). A highly purified 27-kDa protein also has been used as a vaccine in guinea pigs, mice and cattle (17). Recently, Hendrix et al (4,5) reported cloning and sequencing of a gene, corn] , encoding a 27-kDa OMP of the Nine Mile strain and indicated this protein was expressed in both acute and chronic Q fever isolates.…”

mentioning

confidence: 99%

Differentiation of Coxiella burnetii by Sequence Analysis of the Gene (com1) Encoding a 27‐kDa Outer Membrane Protein

Zhang

Yamaguchi

et al. 1997

The gene (coml) encoding a 27-kDa outer membrane protein in 21 strains of Coxiella burnetii from a variety of clinical and geographical sources was sequenced for strain differentiation. The coml gene was highly conserved among all the strains tested but there were several differences in nucleotide and deduced amino acid sequences. Based on the coml gene-specific nucleotides and deduced amino acids, the 21 strains were divided into four groups. Group 1 contained 14 strains originating from ticks, cattle and human cases of acute Q fever. Groups 2 and 3 included 2 and 3 strains, respectively, originating from human cases of chronic Q fever. Group 4 contained 2 strains originating from a human case of acute Q fever and a goat with abortion. The results indicated that the strains originating from ticks, cattle and human cases of acute Q fever differed at the molecular level from those of human chronic Q fever. This study suggests that a sequence analysis of the coml gene can be used for strain differentiation of C. burnetii.