Purification of Encephalitozoon cultures contaminated by mycoplasmas by murine intraperitoneal inoculation.
Encephalitozoon species are strict intracellular microsporidia. Cocultures with eukaryotic cell lines can become accidently contaminated by mycoplasmas. We propose a decontamination protocol based on differential cell targeting after intraperitoneal inoculation in mice. Mycoplasma-free microsporidia were isolated from the brains and spleens of inoculated mice 24 h postinoculation by using the centrifugation shell vial system. Identification was confirmed by direct sequencing of PCR-amplified 16S rRNA.
Unité des Rickettsies CNRS UPRES-A 6020, Faculté de Médecine, Université de la Méditerranée, Marseille, France. Didier.Raoult@medecine.univ-mrs.fr
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Ridoux
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Foucault
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Drancourt
D000818:Animals; D001921:Brain / Q000469:parasitology; D018929:Cell Culture Techniques / Q000379:methods*; D002498:Centrifugation; D002522:Chlorocebus aethiops; D016819:Encephalitozoon / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D009174:Mycoplasma / Q000254:growth & development / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013154:Spleen / Q000469:parasitology; D014709:Vero Cells
D012336:RNA, Ribosomal, 16S