Methods of propagating and detecting infectious forms of intracellular pathogenic microorganisms

Abstract

Methods and kits for detecting the presence of infectious forms of intracellular pathogenic protozoa, particularly infectious forms of protozoa in the phylum Microspora and the phylum Apicomplexa, in clinical and environmental samples are provided The methods comprise isolating the protozoa from the test sample; adding an aliquot of the isolated protozoa to the culture medium of a culture of CHO cells, and assaying for the presence of the protozoa within the CHO cells. The kit comprises an antibody that binds to a protein on the infectious form of the protozoa and a sample of CHO cells. Methods of rapidly propagating intracellular pathogenic bacteria and protozoa, particularly protozoa belonging to the phylum Microspora, are also provided. The methods comprise providing a culture of Chinese Hamster Ovary (CHO) cells; adding the protozoa or bacteria to the medium of the cultured cells, and incubating the cells in the presence of the medium for a time sufficient to allow the protozoa or bacteria to infect the cells. Thereafter, the cells are incubated in medium with or without the protozoa or bacteria for a time sufficient for the protozoa or bacterium to replicate within the infected cells. A model system and method for monitoring the effect of candidate pharmacological agents on the infectivity and development of intracellular protozoa and intracellular bacteria are also provided.

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