Determination of specific DNA of Borrelia burgdorferi in urine of patients with Lyme borreliosis (LB)
Abstract
Objective: Elaboration and verification of a PCR procedure for the detection of Borrelia burgdorferi in the urine of patients with Lyme borreliosis (LB). Methods: A suspension of the resin Chelex 100 was used for the isolation of DNA from urine.
The PCR was designed as a two-step amplification ("nested" PCR). Two sets of primers were used: one for the plasmid gene coding Osp C protein, and the other for chromosonal gene coding 16,168 n rDNNA.
A possible presence of inhibitors causing false negative results was controlled by adding human DNA to the reaction and by co-amplification with human genome primers. Results: Of the 28 patients examined DNA was detected in 20 of them: 1 Ix in neuroborreliosis, 3x in Lyme arthritis, 6x in patients suspected of LB.
Most of the positive results were obtained with the Osp C System (17/20, 85 %), followed by the 16S rDNA system (7/20, 35 %). Amplification with both sets of primers was successful in 4 patients only (4/20, 20 %).
Conclclusion: The feasibility of detecting DNA of B. burgdorferi in urine was verified. Further, it was confirmed, that in the DNA diagnostics of spircpchetes at least two PCR systems must be used concurrently, a finding observed by other authors as well.