Measurement of Babesia bovis infected red blood cells using flow cytometry

Pongpisid Koonyosying; Somdet Srichairatanakool; Saruda Tiwananthagorn; Nattawooti Sthitmatee

doi:10.1016/j.mimet.2022.106641

Measurement of Babesia bovis infected red blood cells using flow cytometry

J Microbiol Methods. 2023 Jan:204:106641. doi: 10.1016/j.mimet.2022.106641. Epub 2022 Nov 30.

Authors

Pongpisid Koonyosying¹, Somdet Srichairatanakool², Saruda Tiwananthagorn³, Nattawooti Sthitmatee⁴

Affiliations

¹ Graduate School of Veterinary Science, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand; Laboratory of Veterinary Vaccine and Biological Products, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand.
² Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
³ Department of Veterinary Bioscience and Veterinary Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand.
⁴ Laboratory of Veterinary Vaccine and Biological Products, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand; Department of Veterinary Bioscience and Veterinary Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand; Excellence Center in Veterinary Bioscience, Chiang Mai University, Chiang Mai 50100, Thailand. Electronic address: drneaw@gmail.com.

PMID: 36460090
DOI: 10.1016/j.mimet.2022.106641

Abstract

Rapid and accurate tools are needed for high-throughput in vitro antibabesial drug testing. In this study, flow cytometry for the measuring of Babesia bovis in vitro culture, was developed using SYBR Green I and compared against the results of fluorescence-based assay and microscopic assay. A high correlation of measured parasitemia was observed with high R² value (R² = 0.9991) between flow cytometry and microscopic analysis. The degree of antibabesial drug sensitivity against B. bovis determined by flow cytometry was 0.424 ± 0.173 μM. Similar to the results of previously published studies involving fluorescence spectrometry-based assay (0.408 ± 0.011 μM) and microscopy-based assay (0.400 ± 0.017 μM). The outcomes of this present study suggest that flow cytometry assay using SYBR Green I can potentially be useful in determining parasitemia and can serve as a rapid alternative method to antibabesial drug testing.

Keywords: Babesia bovis; Flow cytometry; SYBR Green I.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Babesia bovis*
Erythrocytes
Flow Cytometry / methods
Humans
Parasitemia / diagnosis

Substances

SYBR Green I