Enliven: Microbes and Microbial Techniques

An Improved Wolbachia Surface Protein based ELISA
Author(s): Imran A, Seshu J, Richard C. Tamfu, Chambers JP

Introduction: Wolbachia bacteria are among the most successful and intriguing intracellular bacteria in nature. Bacterial surface proteins often function as antigens and host elicited antibodies constitute a sensitive means of detection. Detection of Wolbachia has relied on PCR amplification but PCR is not always a reliable means of detection due to primer specificity, strain diversity, degree of infection, and/or tissue sampled.

Objective: To develop a specific and sensitive ELISA for detection of host elicited IgG/IgM response against Wolbachia.

Methods: His-tagged 'sensing' recombinant Wolbachia Surface Protein (WSP) was expressed in E. coli, affinity purified, and validated by PAGE and Proteomic analyses. Rabbits were immunized with recombinant WSP and resulting antiserum affinity purified, and used in development of the ELISA.

Results: High titer IgG was detected in rabbits immunized with purified recombinant WSP. Qualitative assessment of 26 human serum samples revealed a mix of IgG and IgM. Sera from a variety but limited number of animals indicated a broad but positive IgG response.

Conclusion: A limited number of human and non-human sera were found to contain elicited anti-WSP antibodies. The ELISA reported here is a sensitive and potentially useful means of detecting elicited IgG/IgM in response to Wolbachia.