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Improving the specificity of 16S rDNA‐based polymerase chain reaction for detecting Borrelia burgdorferi sensu lato‐causative agents of human Lyme disease
Author(s) -
Cyr T.L.,
Jenkins M.C.,
Hall R.D.,
Masters E.J.,
McDonald G.A.
Publication year - 2005
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2005.02539.x
Subject(s) - borrelia burgdorferi , sensu , polymerase chain reaction , lyme disease , biology , microbiology and biotechnology , borrelia , 16s ribosomal rna , virology , spirochaetaceae , bacteria , gene , genetics , zoology , antibody , genus
Aims: 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. Methods and 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. Significance and Impact of the Study: 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.