METHODS FOR PURIFICATION OF RICKETTSIA-PROWAZEKII SEPARATED FROM THE HOST TISSUE - A STEP-BY-STEP COMPARISON

  • 1 August 1989
    • journal article
    • research article
    • Vol. 33  (4) , 361-370
Abstract
Two methods of purification of Rickettsii prowazekij 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% biological 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.