Improved chicken embryo cell culture plaque assay for scrub typhus rickettsiae

Abstract:The simple quantification of viable intracellular bacteria is important for the study of an obligate intracellular bacterium, Orientia tsutsugamushi. We applied a novel monoclonal antibody (M686-13) -specific for intracellular Orientia-to an immunofluorescent antibody (IFA) test for determining antibiotic susceptibility of O. tsutsugamushi. M686-13 did not react with Orientia that was inhibited by doxycycline, although bacterial particles still remained in the cells. This preferential staining of proliferating bacteria made the IFA test rapid and precise. Using this method, we could successfully measure the minimal inhibitory concentration (MIC) of a Korean strain of O. tsutsugamushi to doxycycline and clindamycin. This method may be used in other procedures to evaluate the growth of Orientia.

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Use of the Plaque Assay for Testing the Antibiotic Susceptibility of Intracellular Bacteria

Edouard

Raoult

2013

Future Microbiol.

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The plaque assay was first described for titration of bacterial inoculums and clonal isolation, and was later adapted for testing antibiotics susceptibility and to study virulence factors and motility of bacteria. Over time, the sensitivity and reproducibility of the technique has been improved. Usually, the number of plaques is counted; however, the recent development of informatics tools has stimulated interest in the quantification of plaque size. Owing to this new approach, the plaque assay has been used to characterize the host cell response when infected cells are treated with antimicrobial agents. It was found that statins prevented cell lesions following rickettsial infection; in other studies, some antibiotics were found to cause apoptosis of host cells, suggesting a toxic activity. Here, we present an overview of the plaque assay as it has been used to investigate intracellular bacteria.

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Improved chicken embryo cell culture plaque assay for scrub typhus rickettsiae

Abstract: The plaque technique for three strains of Rickettsia tsutsugamushi in chicken embryo cell cultures was greatly improved by modifying the trypsinizing procedure and employing homologous chicken serum in the overlay medium.

Cited by 3 publications

References 4 publications

Plaque assay for scrub typhus rickettsiae

Plaque assay for scrub typhus rickettsiae

Application of Monoclonal Antibody, Specific for Intracellular Orientia tsutsugamushi, to Immunofluorescent Antibody Test for Determining Antibiotic Susceptibility

Use of the Plaque Assay for Testing the Antibiotic Susceptibility of Intracellular Bacteria

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