Study of Lyme neuroborreliosis using polymerase chain reaction

Lyme borreliosis is a multi-system disorder caused by spirochetes of the genus Borrelia. Diagnosis of Lyme disease remains problematic. To aid in the diagnosis and characterization of disorders associated with neuroborreliosis, nested pairs of oligonucleotide primers were designed to recognize the C-terminal region of Borrelia burgdorferi linear plasmid-encoded major surface protein OspA. The assay was used to evaluate cerebrospinal fluid (CSF) from 48 patients thought to have neuroborreliosis, and the results from this PCR-based diagnostic assay were compared to those from current diagnostic methods. This assay detected B. burgdorferi DNA in the CSF of most patients who had both clinical evidence of neuroborreliosis and evidence of Borrelia-specific antibody production in the CSF (18 of 21 patients), and identified patients with clinical evidence of neuroborreliosis who were missed by other diagnostic methods. The described PCR assay detected B. burgdorferi DNA in zero of 23 patient controls and zero of 83 additional contamination controls. Additionally, the assay detected B. burgdorferi DNA in post-treatment CSF samples from some patients who failed to respond to antibiotic treatment or who subsequently relapsed, making it a useful adjunct to existing diagnostic tests and a potential aid in the assessment of treatment response.; To provide an assay which could recognize isolates from the full range of strains associated with Lyme disease and distinguish between infection caused by pathogens from different borrelial species, four sets of nested primers were designed which target the chromosomal DNA encoding 16S ribosomal RNA of Lyme disease-causing borrelial species. Exploiting either the conservation or variability at specific positions in the 16S rDNA target, primers were designed to amplify DNA from the three spirochete species associated with Lyme disease and to exclude amplification of DNA from closely related non-Lyme disease-causing species. One primer set was designed to recognize all three Lyme disease-causing species and each of the other sets was designed to be species-specific. The Lyme borreliosis universal primer sets detected DNA from all Lyme disease-causing borrelial species tested, but not from any of the bacterial negative control strains.