The isolation of Borrelia burgdorferi spirochetes from clinical material in cell line cultures.
It has been found that B. burgdorferi bacteria multiply in mouse fibroblasts. Mouse fibroblast of the L-929 cell line was inoculated with less than 10 up to 10(4) B. burgdorferi cells and incubated for 2-10 days at 35 degrees C in microaerophilic conditions. Within 2 days, visible growth was observed. The bacteria were present in growth medium and on/in mouse fibroblasts as revealed by the indirect immunofluorescence assay. At the same time, development of vacuolized fibroblastic giant cells was observed. Viable spirochetes were also detected in Eagle's medium from a L-929 fibroblast cell line culture, after approximately 2-5 days of incubation with blood, cerebro-spinal and synovial fluids of Lyme borreliosis patients. The bacteria were present in growth medium and on/in endothelial cells as revealed by the indirect immunofluorescence assay. The establishment of B. burgdorferi culture conditions in cell lines gives us a possibility to isolate the etiological agent of Lyme disease from patient blood, cerebrospinal and synovial fluids at different stages of infection. The high sensitivity of this procedure would be helpful in a proper identification of the infection as well as in the control of treatment effectiveness.
Tylewska-Wierzbanowska
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Department of Bacteriology, National Institute of Hygiene, Warsaw, Poland.
Chmielewski
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T
D000818:Animals; D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D002478:Cells, Cultured; D004269:DNA, Bacterial / Q000302:isolation & purification; D004727:Endothelium / Q000382:microbiology; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007739:L Cells; D008193:Lyme Disease / Q000097:blood / Q000134:cerebrospinal fluid / Q000382:microbiology*; D051379:Mice; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013582:Synovial Fluid / Q000382:microbiology
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S