Methods for purification of Rickettsia prowazekii separated from the host tissue: a step-by-step comparison

Two methods for purification of Rickettsia prowazekii strains E, E Vir, and Breinl grown in chick embryo yolk sacs are described. These methods combine either differential centrifugation or sucrose mix, centrifugation through sucrose cushion, 10 mmol/l MgCl2 treatment, filtration through a glass fil...

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Bibliographic Details
Published inActa virologica (Anglickâa verze) Vol. 33; no. 4; p. 361
Main Authors Aniskovich, L P, Eremeeva, M E, Balaeva, N M, Ignatovich, V F, Artemiev, M I, Emelyanov, V V, Smirnova, N S
Format Journal Article
LanguageEnglish
Published Slovakia 01.08.1989
Subjects
Animals
Bacteriological Techniques
Centrifugation, Density Gradient
Chick Embryo
Diatrizoate Meglumine
Filtration
Microscopy, Electron
Rickettsia prowazekii - isolation & purification
Rickettsia prowazekii - ultrastructure
Sucrose
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Summary:Two methods for purification of Rickettsia prowazekii strains E, E Vir, and Breinl grown in chick embryo yolk sacs are described. These methods combine either differential centrifugation or sucrose mix, centrifugation through sucrose cushion, 10 mmol/l MgCl2 treatment, filtration through a glass filter AP-20 and 2 cycles of verografin discontinuous density gradient centrifugation. The purification procedure including sucrose mix allowed to recover about 38-42% biologically active rickettsiae, a yield which was by 10% higher than that obtained by the method beginning at differential centrifugation. The rickettsiae free of host cell components preserved their infectious activity. The obtained biomass was suitable for immunological and biological characterization of Rickettsia prowazekii and for isolation of its total DNA.
ISSN:0001-723X
1336-2305