IDENTIFICATION OF RICKETTSIA-PROWAZEKII USING THE POLYMERASE CHAIN-REACTION

Polymerase chain reaction (PCR) was used to identify Rickettsia prowaz ekii, the etiologic agent of epidemic typhus. For the PCR, Thermus the rmophilus thermostable DNA polymerase was applied with buffer containi ng a relatively low Mg2+ concentration (1.5-2 mM with dNTP's at 250 mu M each). A primer pair used to amplify a 448-base-pair (bp) fragment o f R. prowazekii genome was synthesized on the basis of the DNA sequenc e of gene rpa14/16, coding for a precursor of the mature polypeptides of molecular weight (Mr) 14,000 and/or 16,000 (16kD) from R. prowazeki i strain E. For determining the specificity of the primer pair, purifi ed genomic DNAs of 16 rickettsial and 10 other bacterial strains were used.