Improved Quantification, Propagation, Purification and Storage of the Obligate Intracellular Human Pathogen Orientia tsutsugamushi

PLoS Neglected Tropical Diseases
Suparat GiengkamJeanne Salje

Abstract

Scrub typhus is a leading cause of serious febrile illness in rural Southeast Asia. The causative agent, Orientia tsutsugamushi, is an obligate intracellular bacterium that is transmitted to humans by the bite of a Leptotrombidium mite. Research into the basic mechanisms of cell biology and pathogenicity of O. tsutsugamushi has lagged behind that of other important human pathogens. One reason for this is that O. tsutsugamushi is an obligate intracellular bacterium that can only be cultured in mammalian cells and that requires specific methodologies for propagation and analysis. Here, we have performed a body of work designed to improve methods for quantification, propagation, purification and long-term storage of this important but neglected human pathogen. These results will be useful to other researchers working on O. tsutsugamushi and also other obligate intracellular pathogens such as those in the Rickettsiales and Chlamydiales families. A clinical isolate of O. tsutsugamushi was grown in cultured mouse embryonic fibroblast (L929) cells. Bacterial growth was measured using an O. tsutsugamushi-specific qPCR assay. Conditions leading to improvements in viability and growth were monitored in terms of the effect on bacterial ce...Continue Reading

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Citations

May 18, 2017·Molecular Microbiology·Sharanjeet AtwalJeanne Salje
Nov 4, 2017·PLoS Neglected Tropical Diseases·Guang XuChristine M Arcari
Feb 13, 2018·The Journal of Immunology : Official Journal of the American Association of Immunologists·Lynn Soong
Mar 28, 2021·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Ivo ElliottElizabeth M Batty
Sep 14, 2021·Pathogens and Disease·Jason R Hunt, Jason A Carlyon

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Methods Mentioned

BETA
scraping
PCR
fluorescence
confocal microscopy
light microscopy
fluorescence microscopy

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