A two year prospective study to compare culture and polymerase chain reaction amplification for the detection and diagnosis of Lyme borreliosis.

Picken, Maria M.; Picken, Roger N.; Han, Dong; Cheng, Yu; Ružić‐Sabljić, Eva; Cimperman, J; Maraspin, Vera; Lotrič‐Furlan, Stanka; Strle, Franc

doi:10.1136/mp.50.4.186

Cited by 58 publications

(51 citation statements)

References 30 publications

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“…We contend that it is unwise to draw conclusions about gene regulation from microarray data displaying such relatively small changes. This is particularly true for B. burgdorferi, which is sensitive to many as yet unknown culture variables (Callister et al, 1990;Nelson et al, 1991;Picken et al, 1997;Wang et al, 2004;Yang et al, 2001).…”

Section: Genes Potentially Regulated By Rrp2 Onlymentioning

confidence: 99%

Transcriptional interplay among the regulators Rrp2, RpoN and RpoS in Borrelia burgdorferi

2008

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The RpoN-RpoS alternative sigma factor pathway is essential for key adaptive responses by Borrelia burgdorferi, particularly those involved in the infection of a mammalian host. A putative response regulator, Rrp2, is ostensibly required for activation of the RpoN-dependent transcription of rpoS. However, questions remain regarding the extent to which the three major constituents of this pathway (Rrp2, RpoN and RpoS) act interdependently. To assess the functional interplay between Rrp2, RpoN and RpoS, we employed microarray analyses to compare gene expression levels in rrp2, rpoN and rpoS mutants of parental strain 297. We identified 98 genes that were similarly regulated by Rrp2, RpoN and RpoS, and an additional 47 genes were determined to be likely regulated by this pathway. The substantial overlap between genes regulated by RpoS and RpoN provides compelling evidence that these two alternative sigma factors form a congruous pathway and that RpoN regulates B. burgdorferi gene expression through RpoS. Although several known B. burgdorferi virulence determinants were regulated by the RpoN-RpoS pathway, a defined function has yet to be ascribed to most of the genes substantially regulated by Rrp2, RpoN and RpoS.

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Section: Genes Potentially Regulated By Rrp2 Onlymentioning

confidence: 99%

Transcriptional interplay among the regulators Rrp2, RpoN and RpoS in Borrelia burgdorferi

2008

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“…Although a number of as low as one spirochete can be recovered on culture using laboratoryadapted and continuously propagated strains of B. burgdorferi sensu stricto (17,254,262,331), the sensitivity of culture for clinical specimens is undefined. Compared to PCR for detection of spirochetes in cutaneous specimens, culture has proven to be slightly more sensitive in some studies but not in others (31,165,209,227,256,303,327,380). Such disparate results are likely to be attributable to differences in the various studies in the PCR protocols employed, including type of PCR and/or primer and target selection and/or method of tissue preservation, as well as differences in culture techniques, including size of the skin biopsy sample cultured and/or choice of culture medium.…”

Section: Laboratory Diagnosismentioning

confidence: 99%

“…B. burgdorferi sensu lato can be recovered from various tissues and body fluids of patients with LB, including biopsy (14,25,31,140,165,178,200,204,209,214,221,227,237,256,267,303,327,333,342,369) and lavage (369) specimens of EM skin lesions, biopsy specimens of ACA skin lesions (14,258,267,342), biopsy specimens of borrelial lymphocytoma skin lesions (188), cerebrospinal fluid specimens (60,147,237,267,342), and blood specimens (13,22,189,216,221,322,367,368,374). Anecdotally, recovery of B. burgdorferi sensu lato from other tissue or fluid specimens (189), such as synovial fluid (289), cardiac tissue (312), and iris (265), has also been reported.…”

Section: Laboratory Diagnosismentioning

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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“…Isolation of the etiological agent from patient material is the most reliable method for the diagnosis of borrelial infection (1,14,16,23,26,27,47). In addition, it also provides live microorganisms; data obtained from their further characterization provide potentially valuable information on the geographical distribution, epidemiology, and pathogenesis of borrelial infection and Lyme borreliosis.…”

mentioning

confidence: 99%

Comparison of Borrelia burgdorferi Sensu Lato Strains Isolated from Specimens Obtained Simultaneously from Two Different Sites of Infection in Individual Patients

et al. 2005

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The aim of the present study was to analyze and compare Borrelia strains isolated from two different specimens obtained simultaneously from individual patients with Lyme borreliosis. Fifty such patients and 50 corresponding pairs of Borrelia isolates (100 low-propagated strains) were subjected to genotypic and phenotypic analysis, including pulsed-field gel electrophoresis for species identification and plasmid profile determination and protein profile electrophoresis for the assessment of the presence and molecular masses of separated proteins. The strains were isolated from two distinct skin lesions (12 patients), skin and blood (28 patients), skin and cerebrospinal fluid (8 patients), and blood and cerebrospinal fluid (2 patients). Out of 100 isolates, 63 were typed as B. afzelii and 37 as B. garinii. From each individual specimen only a single Borrelia species was cultured. Comparison of 50 Borrelia strain pairs isolated from two different specimens of an individual patient revealed that 12/50 (24%) patients were simultaneously infected with two different Borrelia strains; in 3/50 (6%) patients strains differed at the species level, in 4 out of the remaining 47 (9%) patients a strain difference in plasmid profile was established, while 5 out of the remaining 43 (11%) patient strain pairs differed in regard to the protein profiles of the two concurrently isolated strains. The results of the present study indicate that human patients with Lyme borreliosis may simultaneously harbor different B. burgdorferi sensu lato strains.

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A two year prospective study to compare culture and polymerase chain reaction amplification for the detection and diagnosis of Lyme borreliosis.

Cited by 58 publications

References 30 publications

Transcriptional interplay among the regulators Rrp2, RpoN and RpoS in Borrelia burgdorferi

Transcriptional interplay among the regulators Rrp2, RpoN and RpoS in Borrelia burgdorferi

Diagnosis of Lyme Borreliosis

Comparison of Borrelia burgdorferi Sensu Lato Strains Isolated from Specimens Obtained Simultaneously from Two Different Sites of Infection in Individual Patients

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