title: Identification of Rickettsia prowazekii using the polymerase chain reaction.
abstract: Polymerase chain reaction (PCR) was used to identify Rickettsia prowazekii, the etiologic agent of epidemic typhus. For the PCR, Thermus thermophilus thermostable DNA polymerase was applied with buffer containing a relatively low Mg2+ concentration (1.5-2 mM with dNTP's at 250 microM each). A primer pair used to amplify a 448-base-pair (bp) fragment of R. prowazekii genome was synthesized on the basis of the DNA sequence 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. prowazekii strain E. For determining the specificity of the primer pair, purified genomic DNAs of 16 rickettsial and 10 other bacterial strains were used.
authors.affiliation: N.F. Gamaleya Research Institute of Epidemiology and Microbiology, Russian Academy of Medical Sciences, Moscow.
authors.forename: L P
authors.initials: LP
authors.lastname: Aniskovich
authors.forename: V L
authors.initials: VL
authors.lastname: Motin
authors.forename: L J
authors.initials: LJ
authors.lastname: Lichoded
authors.forename: N M
authors.initials: NM
authors.lastname: Balayeva
authors.forename: G B
authors.initials: GB
authors.lastname: Smirnov
mesh_terms: D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002642:Chick Embryo; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D004587:Electrophoresis, Agar Gel; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012283:Rickettsia prowazekii / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial