Improving the specificity of 16S rDNA-based polymerase chain reaction for detecting Borrelia burgdorferi sensu lato-causative agents of human Lyme disease.
OBJECTIVE
16S rDNA sequences of Borrelia burgdorferi sensu lato were aligned with the 16S rDNA sequences of Borrelia hermsii, Borrelia turicatae, and Borrelia lonestari in order to identify primers that might be used to more specifically identify agents of human Lyme disease in ticks in human skin samples.


RESULTS
Standard polymerase chain reaction (PCR), using an oligonucleotide sequence, designated TEC1, was shown, in combination with a previously developed primer (LD2) to amplify strains of B. burgdorferi sensu stricto, Borrelia afzelii, and Borrelia garinii, but not the non-Lyme causing B. hermsii or B. turicatae. This primer pair, designated Bbsl, was successfully used to amplify B. burgdorferi sensu lato from skin biopsies of patients with Lyme disease symptoms as well as from Ixodes scapularis, Amblyomma americanum and Dermacentor variabilis ticks.


CONCLUSIONS
The primer set Bbsl allows for the rapid detection and differentiation of B. burgdorferi sensu lato from non-Lyme disease-causing Borrelia species in ticks and human tissues.


CONCLUSIONS
The PCR primer set, Bbsl, will greatly facilitate detection of the causative agents of Lyme disease in infected ticks and human skin samples assisting in epidemiological studies, and potentially allowing for a more rapid diagnosis of the disease in patients.
Cyr
T L
TL
Animal Parasitic Diseases Laboratory, Agricultural Research Service, USDA-ARS, Building 1040, Room 100, BARC-East, Beltsville, MD 20705, USA. tcyr@anri.barc.usda.gov
Jenkins
M C
MC
Hall
R D
RD
Masters
E J
EJ
McDonald
G A
GA
D000328:Adult; D000368:Aged; D000818:Animals; D001096:Arachnid Vectors / Q000235:genetics; D025065:Borrelia burgdorferi / Q000235:genetics; D015748:Borrelia burgdorferi Group / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis*; D004275:DNA, Ribosomal / Q000235:genetics*; D003870:Dermacentor / Q000235:genetics; D005260:Female; D005798:Genes, Bacterial / Q000235:genetics; D006801:Humans; D018884:Ixodes / Q000235:genetics; D008193:Lyme Disease / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA / Q000379:methods; D012867:Skin / Q000473:pathology; D013987:Ticks / Q000235:genetics*
D004269:DNA, Bacterial; D004275:DNA, Ribosomal