Antibody response to Brucella outer membrane proteins in bovine brucellosis: immunoblot analysis and competitive enzyme-linked immunosorbent assay using monoclonal antibodies.

AbstractLeptospiral outer membrane proteins (OMPs) are highly conserved in different species, and play an essential role in the development of new immunoprotection and serodiagnosis strategies. The genes encoding LipL21, LipL32 and OmpL1 were cloned from the complete genome sequence of Leptospira interrogans serovar lai strain Lai and expressed in vitro. Sequence comparison analysis revealed that the three genes were highly conserved among distinct epidemic leptospires, including three major epidemic species Leptospira interrogans, Leptospira borgpetersenii and Leptospira weilii, in China. Immunoblot analysis was further performed to scrutinize 15 epidemic Leptospira reference strains using the antisera of the recombinant OMPs. Both immunoblot assay and reverse transcription-polymerase chain reaction demonstrated that these three OMPs were conservatively expressed in pathogenic L. interrogans strains and other pathogenic leptospires. Additionally, the use of these recombinant OMPs as antigens in enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of leptospirosis was evaluated. The recombinant LipL32 and OmpL1 proteins showed a high degree of ELISA reactivity with sera from patients infected with L. interrogans strain Lai and other pathogenic leptospires. These results may contribute to the identification of candidates for broad-range vaccines and immunodiagnostic antigens in further research.

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Serological diagnostic potential of recombinant outer membrane proteins (rOMPs) from Brucella melitensis in mouse model using indirect enzyme-linked immunosorbent assay

BMC Veterinary Research ◽

10.1186/s12917-015-0587-2 ◽

2015 ◽

Vol 11 (1) ◽

Cited By ~ 8

Author(s):

Ihsan Muneer Ahmed ◽

Siti Khairani-Bejo ◽

Latiffah Hassan ◽

Abdul Rani Bahaman ◽

Abdul Rahman Omar

Keyword(s):

Membrane Proteins ◽

Mouse Model ◽

Outer Membrane ◽

Immunosorbent Assay ◽

Brucella Melitensis ◽

Outer Membrane Proteins ◽

Enzyme Linked Immunosorbent Assay ◽

Diagnostic Potential

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An Enzyme-Linked Immunosorbent Assay for the Rapid Detection of Vibrio parahaemolyticus

Journal of Food Protection ◽

10.4315/0362-028x-58.8.873 ◽

1995 ◽

Vol 58 (8) ◽

pp. 873-878 ◽

Cited By ~ 8

Author(s):

CHI H. CHEN ◽

TSUNG C. CHANG

Keyword(s):

Membrane Proteins ◽

Outer Membrane ◽

Immunosorbent Assay ◽

Vibrio Parahaemolyticus ◽

Rapid Detection ◽

Outer Membrane Proteins ◽

Screening Method ◽

Rapid Screening ◽

Enzyme Linked Immunosorbent Assay ◽

Conventional Culture

A sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the rapid detection of Vibrio parahaemolyticus in foods. The assay was based on the detection of two outer-membrane proteins (molecular weights 36,000 and 34,000) of V. parahaemolyticus. Following an 18-h incubation in alkaline peptone salt broth containing 0.1%Teepol, the culture supernatant was added to the microtiter plate coated with antibodies against the two outer-membrane proteins. After washing, the same antibodies absorbed with V. alginolyticus and labeled with horsemdish peroxidase were used as secondary antibodies. The detection limit of the assay for total outer-membrane proteins was 10 ng/ml. For 29 strains of V. parahaemolyticus and 73 strains (including 27 isolates of vibrios) of other bacteria tested, the sensitivity and specificity of the ELISA were 100 and 96%, respectively. Strains producing false positives were V. tubiashii, V. campbellii, and V. vulnificus. Of 23 seafood samples tested, V. parahaemolyticus was detected in 17 and 15 samples, respectively, by the ELISA and by a conventional culture method. V. parahaemolyticus was also detected in samples artificially inoculated with the microorganism at levels less than 10 colony-forming units (CFU) per g. Compared to the conventional culture methods, which may take 4 to 6 days to complete, the ELISA can detect low numbers of V.parahaemolyticus in foods with a total analytical time of only 24 h. The ELISA is recommended as a rapid screening method.

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