The Polymerase Chain Reaction (PCR) to Detect Gene Sequences of Borrelia Burgdorferi, the Etiologic Agent of Lyme Disease

Abstract

The direct demonstration of Borrelia burgdorferi (B. burgdorferi) in specimens from infected tissues is difficult because only very few spirochetes appear to be present. Since the polymerase chain reaction (PCR) can be used to detect small numbers of organisms it can be envisaged as an alternative diagnostic method for pathogen detection. Since it was important to select DNA target sequences that are highly conserved among different isolates of B. burgdorferi we have chosen as target DNA sequence the gene encoding flagellin, a structure that had been found to show only minimal variation within the B. burgdorferi species. Various combinations of different oligonucleotide primers were tested for their efficiency. The most efficient primer combination allowed to detect either 0.2 pg of B. burgorferi DNA by visual examination of ethidium bromide stained gels and of 0.01 pg DNA after radioactive hybridization.