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Development and optimization of an in vitro cultivation protocol allows for isolation of Borrelia miyamotoi from patients with hard tick-borne relapsing fever J. Koetsveld, N.M. Kolyasnikova, A. Wagemakers, M.G. Toporkova, D.S. Sarksyan, A. Oei, A.E. Platonov, J.W. Hovius Clinical Microbiology and Infection Volume 23, Issue 7, Pages (July 2017) DOI: /j.cmi Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases Terms and Conditions
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Fig. 1 The influence of anticoagulants on Borrelia miyamotoi culture and our protocol for culturing B. miyamotoi from human blood. (a) A serial dilution of B. miyamotoi (strain HT31) spirochaetes was prepared and 1 to 1000 spirochaetes were directly inoculated to fresh MKP-F and used for in vitro culture during 1 month. The combined data of three different experiments are shown; bars and error bars represent the mean±standard deviation. (b) Serial dilutions of lithium heparin (0.25–5 IU/mL), K2-EDTA (25–500 μg/mL) and trisodium citrate (0.25–5 mM) were prepared in MKP-F. A total of 1000 B. miyamotoi (strain HT31) spirochaetes were added to each condition. Spirochaete concentrations that were reached after 1 month of cultivation are shown; bars and error bars represent the mean±standard deviation. (c) Heparin blood from three healthy blood donors was spiked with 1 to 1×104 B. miyamotoi strain HT31 spirochaetes per mL blood. A total of 1 mL of plasma was collected and centrifuged at 8000 g for 8 min. The pellet was resuspended in MKP-F and added to 7 mL fresh MKP-F culture tubes. Bars and error bars represent the mean±standard deviation. Clinical Microbiology and Infection , DOI: ( /j.cmi ) Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases Terms and Conditions
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Fig. 2 Cultivation protocol for Borrelia miyamotoi isolation from human blood. A two-step centrifugation protocol for isolation of B. miyamotoi from blood of patients with hard tick-borne relapsing fever. Clinical Microbiology and Infection , DOI: ( /j.cmi ) Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases Terms and Conditions
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Fig. S1 Time-to-kill assay.
Clinical Microbiology and Infection , DOI: ( /j.cmi ) Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases Terms and Conditions
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