Literature DB >> 15068385

Improvement in the laboratory recognition of lyme borreliosis with the combination of culture and PCR methods.

Tomasz Chmielewski1, Janusz Fiett, Marek Gniadkowski, Stanislawa Tylewska-Wierzbanowska.   

Abstract

BACKGROUND: Lyme disease is a multisystem, multistage infection caused by three genospecies of the Borrelia burgdorferi sensu lato species. The diagnosis of Lyme disease is based on a history of tick-bite, physical examination, and serological tests. In the seronegative patients with Lyme borreliosis symptoms, additional testing should be introduced.
METHODS: The study group was composed of 240 hospitalized patients presented with various clinical symptoms suggesting Lyme borreliosis: 221 of the patients with neurological abnormalities and 19 with oligoarticular arthritis. Citrated blood and serum samples were collected from the patients for culture and serological examination, respectively. Moreover, 173 cerebrospinal and 6 synovial fluid samples were tested. New oligonucleotide primers based on B. burgdorferi sensu lato 16SrRNA gene sequences were designed for the detection of the bacteria in blood, cerebrospinal, and synovial fluid specimens with PCR. Levels of specific antibodies were measured in serum, cerebrospinal fluid and synovial fluid samples using ELISA and Western blot. B. burgdorferi spirochetes from blood, cerebrospinal fluid, and synovial fluid samples were cultured in cell line. Extracted and purified B. burgdorferi DNA was identified by PCR with new oligonucleotide primers. Then three genospecies were identified by PCR amplification with other primer sets specific for 16S rDNA and/or by the restriction fragment length polymorphism of 23S(rrl)-5S(rrf).
RESULTS: Bacterial DNA were found in samples from 32 patients, including 28 patients with neuroborreliosis and 4 with Lyme arthritis. B. burgdorferi-specific IgM and/or IgG serum antibodies were detected in 14 of these patients. Fourteen strains of Borrelia garini, 4 strains of Borrelia afzelii and 1 strain of B. burgdorferi sensu stricto were identified by PCR. Genospecies were not recognized in 13 specimens.
CONCLUSIONS: The procedure can be a rapid and sensitive diagnostic method for the detection of etiological agents in clinical materials derived from patients with the clinical symptoms of Lyme borreliosis. It can be utilized for both basic research as well as routine laboratory diagnosis.

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Year:  2003        PMID: 15068385     DOI: 10.1007/bf03260032

Source DB:  PubMed          Journal:  Mol Diagn        ISSN: 1084-8592


  31 in total

1.  Development of polymerase chain reaction primer sets for diagnosis of Lyme disease and for species-specific identification of Lyme disease isolates by 16S rRNA signature nucleotide analysis.

Authors:  R T Marconi; C F Garon
Journal:  J Clin Microbiol       Date:  1992-11       Impact factor: 5.948

2.  Identification of a third genomic group of Borrelia burgdorferi through signature nucleotide analysis and 16S rRNA sequence determination.

Authors:  R T Marconi; C F Garon
Journal:  J Gen Microbiol       Date:  1992-03

3.  Detecting Borrelia burgdorferi in blood from patients with Lyme disease.

Authors:  R B Nadelman; I Schwartz; G P Wormser
Journal:  J Infect Dis       Date:  1994-06       Impact factor: 5.226

4.  Detection of Borrelia burgdorferi DNA by polymerase chain reaction in cerebrospinal fluid in Lyme neuroborreliosis.

Authors:  J J Nocton; B J Bloom; B J Rutledge; D H Persing; E L Logigian; C H Schmid; A C Steere
Journal:  J Infect Dis       Date:  1996-09       Impact factor: 5.226

5.  Use of serum immune complexes in a new test that accurately confirms early Lyme disease and active infection with Borrelia burgdorferi.

Authors:  M Brunner; L H Sigal
Journal:  J Clin Microbiol       Date:  2001-09       Impact factor: 5.948

6.  The isolation of Borrelia burgdorferi spirochetes from clinical material in cell line cultures.

Authors:  S Tylewska-Wierzbanowska; T Chmielewski
Journal:  Zentralbl Bakteriol       Date:  1997-10

7.  An OspA serotyping system for Borrelia burgdorferi based on reactivity with monoclonal antibodies and OspA sequence analysis.

Authors:  B Wilske; V Preac-Mursic; U B Göbel; B Graf; S Jauris; E Soutschek; E Schwab; G Zumstein
Journal:  J Clin Microbiol       Date:  1993-02       Impact factor: 5.948

8.  Phylogenetic analysis of the genus Borrelia: a comparison of North American and European isolates of Borrelia burgdorferi.

Authors:  R T Marconi; C F Garon
Journal:  J Bacteriol       Date:  1992-01       Impact factor: 3.490

9.  PCR detection of Borrelia burgdorferi DNA in cerebrospinal fluid of Lyme neuroborreliosis patients.

Authors:  T L Keller; J J Halperin; M Whitman
Journal:  Neurology       Date:  1992-01       Impact factor: 9.910

10.  Changes in infectivity and plasmid profile of the Lyme disease spirochete, Borrelia burgdorferi, as a result of in vitro cultivation.

Authors:  T G Schwan; W Burgdorfer; C F Garon
Journal:  Infect Immun       Date:  1988-08       Impact factor: 3.441

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  3 in total

1.  Ankle arthritis in a 6-year-old boy after a tick bite - a case report.

Authors:  Per-Henrik Randsborg; Carl-Erik Naess
Journal:  Open Orthop J       Date:  2011-05-02

2.  Bacterial tick-borne diseases caused by Bartonella spp., Borrelia burgdorferi sensu lato, Coxiella burnetii, and Rickettsia spp. among patients with cataract surgery.

Authors:  Tomasz Chmielewski; Joanna Brydak-Godowska; Beata Fiecek; Urszula Rorot; Elżbieta Sędrowicz; Małgorzata Werenowska; Dorota Kopacz; Agata Hevelke; Magdalena Michniewicz; Dariusz Kęcik; Stanisława Tylewska-Wierzbanowska
Journal:  Med Sci Monit       Date:  2014-06-05

3.  Empirical validation of the Horowitz Multiple Systemic Infectious Disease Syndrome Questionnaire for suspected Lyme disease.

Authors:  Maryalice Citera; Phyllis R Freeman; Richard I Horowitz
Journal:  Int J Gen Med       Date:  2017-09-04
  3 in total

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