Evaluation of Indigenous latex agglutination assay based on recombinant pasteurella lipoprotein E (rPlpE) as antigen for detection of anti Mannheimia haemolytica - IgG Antibodies

Document Type : Original Articles

Authors

1 Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

2 Department of Pathobiology, Faculty of Veterinary Medicine, University of Shahrekord, Shahrekord, Iran

10.22092/ari.2024.364038.2934

Abstract

Mannheimia haemolytica (M. haemolytica) is an organism causing pneumonia in ruminants. M. haemolytica leads to severe economic losses to the global livestock industry. Different diagnostic methods have been developed such as bacterial culture, bacterial DNA detection and serological assays. Diagnosis of M. haemolytica is based on the bacteriological methods such as isolation of the microorganisms from clinical specimens. Available methods are time-consuming and not easy to perform. Serological tests based on recombinant protein may provide higher sensitivity and specificity than culture tests. There is a need for new diagnostic methods for detection of M.haemolytica specific antibodies. Therefore, it is important to develop an accurate methods for rapid detection of M. haemolytica. In this study, latex agglutination test (LAT) was developed based on recombinant outer membrane pasteurella lipoprotein E (rPlpE) for detecting specific antibodies against M. haemolytica. Recombinant PlpE was constructed using pET26 (b) expression vector in Escherichia coli. Expressed recombinant PlpE was purified and coated with latex particles for a latex agglutination test. The recombinant PlpE was able to detect anti-M. haemolytica IgG in positive sera but did not show any immunoreaction with Pasteurella multocida and negative samples. These results suggest that the rPlpE can be used to detect the specific anti Mannheimia haemolytica - IgG Antibodies. Because the recombinant proteins can be produced efficiently and are cost-effective, their use in diagnostic kits such as LATs as reagents can reduce the cost of them. This rapid and specific anti M. haemolytica antibodies detection method using recombinant proteins can save cost and be widely applied for efficient and practical detection of. M. haemolytica.

Keywords: Antibody, Latex agglutination test, Mannheimia haemolytica, PlpE, Recombinant protein

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Archives of Razi Institute

Articles in Press, Accepted Manuscript
Available Online from 24 February 2024

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