A NESTED POLYMERASE CHAIN-REACTION FOR THE DETECTION OF BORRELIA-BURGDORFERI SENSU-LATO BASED ON A MULTIPLE SEQUENCE-ANALYSIS OF THE HBB GENE

A highly sensitive nested polymerase chain reaction method was designe d for the detection of a wide spectrum of strains from Borrelia burgdo rferi sensu late. This technique allows the detection of as little as 3 fg of total genomic DNA extracted and purified from pure cultures of the organism, this amount corresponds to less than 10 organisms. Two sets of primers homologous to conserved spots in the coding region of the hbb gene, encoding a conserved histone-like protein, were construc ted. These were based on a multiple sequence alignment of 39 strains r epresenting all the genomic groups described in B. burgdorferi sensu l ate.