Polymerase chain reaction for detection ofBorrelia burgdorferi DNA in skin lesions of early and late Lyme borreliosis

Stedingk, Lars‐Victor von; Olssøn, Ingrid; H, Hanson; Asbrink, E; Hovmark, A

doi:10.1007/bf02112610

Cited by 64 publications

(42 citation statements)

References 22 publications

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“…PCR targets that have been employed for detection of B. burgdorferi sensu lato DNA in skin biopsy specimens include p66 (31,210,211,344,358,359), the 16S rRNA gene (296,303), fla (177,234,256), the 23S rRNA gene (31), the 5S rRNA-23S rRNA gene spacer (279), recA (177), and ospA (209,279,327).…”

Section: Vol 18 2005 Diagnosis Of Lyme Borreliosis 489mentioning

confidence: 99%

Diagnosis of Lyme Borreliosis

et al. 2005

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A large amount of knowledge has been acquired since the original descriptions of Lyme borreliosis (LB) and of its causative agent, Borrelia burgdorferi sensu stricto. The complexity of the organism and the variations in the clinical manifestations of LB caused by the different B. burgdorferi sensu lato species were not then anticipated. Considerable improvement has been achieved in detection of B. burgdorferi sensu lato by culture, particularly of blood specimens during early stages of disease. Culturing plasma and increasing the volume of material cultured have accomplished this. Further improvements might be obtained if molecular methods are used for detection of growth in culture and if culture methods are automated. Unfortunately, culture is insensitive in extracutaneous manifestations of LB. PCR and culture have high sensitivity on skin samples of patients with EM whose diagnosis is based mostly on clinical recognition of the lesion. PCR on material obtained from extracutaneous sites is in general of low sensitivity, with the exception of synovial fluid. PCR on synovial fluid has shown a sensitivity of up to >90% (when using four different primer sets) in patients with untreated or partially treated Lyme arthritis, making it a helpful confirmatory test in these patients. Currently, the best use of PCR is for confirmation of the clinical diagnosis of suspected Lyme arthritis in patients who are IgG immunoblot positive. PCR should not be used as the sole laboratory modality to support a clinical diagnosis of extracutaneous LB. PCR positivity in seronegative patients suspected of having late manifestations of LB most likely represents a false-positive result. Because of difficulties in direct methods of detection, laboratory tests currently in use are mainly those detecting antibodies to B. burgdorferi sensu lato. Tests used to detect antibodies to B. burgdorferi sensu lato have evolved from the initial formats as more knowledge on the immunodominant antigens has been collected. The recommendation for two-tier testing was an attempt to standardize testing and improve specificity in the United States. First-tier assays using whole-cell sonicates of B. burgdorferi sensu lato need to be standardized in terms of antigen composition and detection threshold of specific immunoglobulin classes. The search for improved serologic tests has stimulated the development of recombinant protein antigens and the synthesis of specific peptides from immunodominant antigens. The use of these materials alone or in combination as the source of antigen in a single-tier immunoassay may someday replace the currently recommended two-tier testing strategy. Evaluation of these assays is currently being done, and there is evidence that certain of these antigens may be broadly cross-reactive with the B. burgdorferi sensu lato species causing LB in Europe

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Section: Vol 18 2005 Diagnosis Of Lyme Borreliosis 489mentioning

confidence: 99%

Diagnosis of Lyme Borreliosis

et al. 2005

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“…ACA is a late dermatologic manifestation of Lyme borreliosis; nevertheless, the PCR sensitivity (61 to 91%) in skin specimens is comparable to that in EM patients (114,116,119,151,153,179). Surprisingly, in four of five patients with histologically proven disease, B. burgdorferi DNA could also be found in urine specimens (155).…”

Section: Patients With Dermatological Manifestationsmentioning

confidence: 91%

“…Correlation of PCR results with clinical improvement could also be shown in patients with late disease, e.g., patients with neuroborreliosis (59,75,84,88,93); patients with ACA, morphea, and lichen sclerosus et atrophicus (1,4,114,155,179); patients with LA (19,22,52,78,123); and a patient with osteomyelitis (126).…”

Section: Monitoring Of Treatment Efficacymentioning

confidence: 96%

“…Recently, it has been shown that one strain isolated in the Czech Republic also failed to amplify with primers specific for the clone 2H1 gene (128). By using clone 2H1 primers and a nested PCR system, 12 (80%) of 15 biopsy samples from nine patients with erythema migrans (EM) or acrodermatitis chronica atrophicans (ACA) were found to be reactive (114); a sensitivity of 69% was found in skin biopsy samples from 26 patients with EM (179). Serum specimens (n ϭ 12), urine specimens (n ϭ 10), and CSF specimens (n ϭ 2) collected from 18 patients with neuroborreliosis were all reactive by PCR with nested primers specific for this chromosomal gene (32).…”

Section: Target Selectionmentioning

confidence: 99%

“…A test sensitivity of 60 to 80% was obtained by using nested PCR with frozen skin tissues (Table 3). Interestingly, positive results were sometimes found only when the DNA preparation was diluted 1:10 or 1:100, indicating an inhibitory effect (179). In a Finnish study, six of seven patients with untreated EM (culture positive) were PCR reactive in a standard PCR with primers specific for the ospA gene (143).…”

Section: Patients With Dermatological Manifestationsmentioning

confidence: 99%

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