disclosed are oligonucleotides useful in methods for determining whether a sample contains borrelia afzelii , a spirochete which is a causative agent of lyme disease in humans . these oligonucleotides , which have nucleotide sequences derived from a coding segment of the gene encoding the p24 gene for the outer surface protein of borrelia afzelii , are useful as forward and reverse primers for a polymerase chain reaction using nucleic acids from a biological sample as templates , and as probes for detecting any resultant amplicon . detection of an amplicon indicates the sample contains borrelia afzelii . real - time pcr and detection using florescence resonance energy transfer is disclosed .