Use of polymerase chain reaction in verification and differential diagnosis of babesiosis pathogens

  • I. I. Torianyk Institute Microbiology and Immunology named after I. I. Mechnikov of the National Academy of Medical Sciences of Ukraine
  • O. M. Tymchenko Institute Microbiology and Immunology named after I. I. Mechnikov of the National Academy of Medical Sciences of Ukraine
  • M. O. Ostapets National University of Pharmacy
  • N. A. Chygyrynska Institute Microbiology and Immunology named after I. I. Mechnikov of the National Academy of Medical Sciences of Ukraine
  • S. I. Pokhyl Institute Microbiology and Immunology named after I. I. Mechnikov of the National Academy of Medical Sciences of Ukraine
  • I. A. Kostyria Institute Microbiology and Immunology named after I. I. Mechnikov of the National Academy of Medical Sciences of Ukraine
  • I. V. Sorokina Kharkiv National Medical University
Keywords: Babesia; verification; blood; homogenates of ticks; cultures of Babesia; polymerase chain reaction.

Abstract

Today, Babesia is recognized as one of the most common blood parasites in the world, which in terms of the number of cases of invasion is second only to trypanosomes (the causative agent of African trypanosomiasis and Chagas’ disease). These microorganisms can cause parasitism in erythrocytes and hematopoietic organs. They cause an infectious process, the clinical course of which can vary from asymptomatic, subclinical, mild or moderate influenza-like forms – to severe progressive disease (fulminant form) with fatal outcome. Thus, the latter determines the significant burden of babesia for the leading branches of medicine, veterinary medicine and the economy as a whole. The presented work is devoted to the study of the prospects for verification of babesiosis causative agents by the polymerase chain reaction (PCR) method. Blood, erythrocyte suspension, homogenized tick-carriers of babesiosis, culture of Babesia spp. were used as research material (samples). In order to obtain an objective assessment, the PCR-diagnostics method was used in two formats – standard and multiplex (multi-primer). Multiple PCR testing of multiplex format using primers in model samples containing cells of different species of Babesia (B. microti, B. divergens, B. bovis, B. canis), allowed us to establish the level of reproducibility of the results of such studies, which ranged 94.6–96.4%, to determine the level of PCR sensitivity of the multiplex format for detection/identification of human pathogenic babesia (B. microti, B. divergens and B. venatorum). It is established that the advantages of the PCR-diagnostic method of babesiosis pathogens in the samples of the studied biomaterial were: speed of research (2–4 hours); high sensitivity, specificity, reproducibility of Babesia detection results, prospects of species identification, differentiation with apicomplex spores (Plasmodium falciparum, Toxoplasma). In view of the above, the PCR method is recommended for use in cases of persistent suspicion of babesiosis infection (in cases of negative results of microscopic/cytological studies, to identify asymptomatic, subclinical and chronic forms of babesiosis, verification of active invasion in seropositive individuals and for Babesia species and their differentiation).

References

Beck, R., Vojta, L., Ćurković, S., & Mrljak, V. (2011). Molecular survey of Babesia microti in wild rodents in Central Croatia. Vector Borne Zoonotic Diseases, 11, 81–83.

Criado-Fornelio, A., Martinez-Marcos, A. Buling-Saraña, A., & Barba-Carretero, J. C. (2003). Presence of Mycoplasma haemofelis, Mycoplasma haemominutum and piroplasmids in cats from southern Europe: A molecular study. Veterinary Microbiology, 93, 307–317.

Ekpanyapong, S., & Reddy, K. R. (2020). Fungal and parasitic infections of the liver. Gastroenterology Clinics of North America, 49(2), 379–410.

Fang, D. C., & McCullough, J. (2016). Transfusion-transmitted Babesia microti. Transfusion Medicine Reviews, 30(3), 132–138.

Gray, J. S., & Weiss, L. M. (2008). Babesia microti. In: Khan, N. (Ed.). Emerging protozoan pathogens. Taylor and Francis, Abingdon. Pp. 303–349.

Gray, J. S., Zintl, A., & Hildebrandt, A. (2010). Zoonotic babesiosis: Overview of the disease and novel aspects of pathogen identity. Ticks and Tick-Borne Diseases, 1, 3–10.

Herwaldt, B. L., Cacciò, S., Gherlinzoni, F., Aspock, H., Slemenda, S. B., Piccaluga, P., Martinelli, G., Edelhofer, R., Hollenstein, U., Poletti G., Pampiglione, S., Loschenberger, K., Tura, S., & Pieniazek, N. J. (2003). Molecular characterization of a non-Babesia divergens organism causing zoonotic babesiosis in Europe. Emerging Infectious Diseases, 9(8), 942–948.

Homer, M., Delfin, J. I. A., & Telford, F. (2000). Babesiosis. Clinical Microbiology Reviewer, 13(3), 451–469.

Kim, A. C., & Epstein, M. E. (2001). Coinfection with Babesia microti and Borrelia burgdorferi. Infection Medicine, 18, 271–276.

Kim, J. Y., Cho, S. H., & Joo, H. N. (2007). First case of human babesiosis in Korea: Detection and characterization of a novel type of Babesia sp. (KO1) similar to ovine Babesia. Journal of Clinical Microbiology, 45(6), 2084–2087.

Kjemtrup, A. M., & Conrad, P. A. (2006). A review of the small canine piroplasms from California: Babesia conradae in the literature. Veterinary Parasitology, 138, 112–117.

Krause Peter, J. (2019). Human babesiosis. International Journal for Parasitology, 49(2), 165–174.

Laha, R., Das, M., & Sen, A. (2015). Morphology, epidemiology, and phylogeny of Babesia: An overview. Symposium, 5(2), 94–100.

Mojtahed, A., Bates, D. D. B., & Hahn, P. F. (2020). Splenic findings in patients with acute babesiosis. Abdominal Radiology, 45, 710–715.

Parija, S. C., Dinoop, K. P., & Venugopal, H. (2015). Diagnosis and management of human babesiosis. Tropical Parasitology, 5(2), 88–93.

Solano-Gallego, L., Sainz, Á., & Roura, X. (2016). A review of canine babesiosis: The European perspective. Parasites Vectors, 9, 336.

Vannier, E., & Krause, P. J. (2012). Human babesiosis. The New England Journal of Medicine, 366(25), 2397–2407.

Zhou, X., Li, S. G., & Chen, S. B. (2013). Co-infections with Babesia microti and Plasmodium parasites along the China-Myanmar border. Infectious Diseases Poverty, 2(1), 24.

Zhou, X., Xia, S., Huang, J. L., Tambo, E., Zhuge, H. X., & Zhou, X. N. (2014). Human babesiosis, an emerging tick-borne disease in the People’s Republic of China. Parasites and Vectors, 7, 509.

Published
2020-11-17
How to Cite
Torianyk, I. I., Tymchenko, O. M., Ostapets, M. O., Chygyrynska, N. A., Pokhyl, S. I., Kostyria, I. A., & Sorokina, I. V. (2020). Use of polymerase chain reaction in verification and differential diagnosis of babesiosis pathogens . Regulatory Mechanisms in Biosystems, 11(4), 563-567. https://doi.org/10.15421/022087