Isolation of Vibrio cholerae O1 and O139 using immunomagnetic separation

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Sahyun Hong, Jong Hyun Kim, Sun Jin Lee, Cheon-Kwon Yoo, Shin-Jung Kang and Gyung Tae Chung

Korea National Institute of Health, Republic of Korea

: J Immunol Tech Infect Dis

Abstract

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Background: Detection of pathogenic microorganisms in environmental samples and patient stools is a difficult process. Objectives: The objective of this study was to develop a method combining immunomagnetic separation (IMS) with PCR or selection media isolation method for a rapid and easily detection of Vibrio cholerae O1 and O139. Methods: Magnetic bead were covalently bound with rabbit anti-V. cholerae O1 or O139 via protein G. This research is an attempt to use magnetic beads for isolating V. cholera O1 and O139 from mixed microorganisms in patient specimen, natural environment and foods. Results and Conclusions: Isolation of V. cholerae O1 and O139 from V. cholera NAG, V. parahaemolyticus, V. vulnificus was examined. In terms of sensitivity and specificity, V. cholerae O1 and O139 1~10CFU/ml couldn’t be isolated but isolation ratio was higher than 95% at 102CFU/ml. The isolation ratio of V. cholerae O1 and O139 from spiked stools was higher than 90% in average. In particular, the PCR was positive in all samples spending for just one hour in IMS method. Such finding demonstrated that an IMS method is effective in isolation V. cholerae O1 and O139 from specimen. This method will reduce laboratory labor, cost, and times.

Biography

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Sahyun Hong Completed Ph.D. in the Department of Biotechnology from Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan. He is a Staff Scientist in the Division of Enteric Bacterial Infections, Center for Infectious Diseases, Korea National Institute of Health, Republic of Korea.

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