Detection of established virulence genes and plasmids to differentiate Borrelia burgdorferi strains

Kamfai Chan, Sherwood Casjens, Nikhat Parveen

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Borrelia burgdorferi sensu stricto is the major causative agent of Lyme disease in the United States, while B. garinii and B. afzelii are more prevalent in Europe. The highly complex genome of B. burgdorferi is comprised of a linear chromosome and a large number of variably sized linear and circular plasmids. Many plasmids of this spirochete are unstable during its culture in vitro. Given that many of the B. burgdorferi virulence factors identified to date are plasmid encoded, spirochetal plasmid content determination is essential for genetic analysis of Lyme pathogenesis. Although PCR-based assays facilitate plasmid profiling of sequenced B. burgdorferi strains, a rapid genetic content determination strategy for nonsequenced strains has not yet been described. In this study, we combined pulsed-field gel electrophoresis (PFGE) and Southern hybridization for detection of genes encoding known virulence factors, ribosomal RNA gene spacer restriction fragment length polymorphism types (RSTs), ospC group determination, and sequencing of the variable dbpA and ospC genes. We show that two strains isolated from the same tick and both originally named N40 are in fact very distinct. Furthermore, we failed to detect bbk32, which encodes a fibronectinbinding adhesin, in one "N40" strain. Thus, two distinct strains that show different plasmid profiles, as determined by PFGE and PCR, were isolated from the same tick and vary in their ospC and dbpA sequences. However, both belong to group RST3B.

Original languageEnglish (US)
Pages (from-to)1519-1529
Number of pages11
JournalInfection and immunity
Volume80
Issue number4
DOIs
StatePublished - Apr 1 2012

Fingerprint

Borrelia burgdorferi
Virulence
Plasmids
Genes
Pulsed Field Gel Electrophoresis
Ticks
Virulence Factors
Polymerase Chain Reaction
Spirochaetales
Lyme Disease
rRNA Genes
Restriction Fragment Length Polymorphisms
Chromosomes
Genome

All Science Journal Classification (ASJC) codes

  • Infectious Diseases
  • Parasitology
  • Microbiology
  • Immunology

Cite this

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abstract = "Borrelia burgdorferi sensu stricto is the major causative agent of Lyme disease in the United States, while B. garinii and B. afzelii are more prevalent in Europe. The highly complex genome of B. burgdorferi is comprised of a linear chromosome and a large number of variably sized linear and circular plasmids. Many plasmids of this spirochete are unstable during its culture in vitro. Given that many of the B. burgdorferi virulence factors identified to date are plasmid encoded, spirochetal plasmid content determination is essential for genetic analysis of Lyme pathogenesis. Although PCR-based assays facilitate plasmid profiling of sequenced B. burgdorferi strains, a rapid genetic content determination strategy for nonsequenced strains has not yet been described. In this study, we combined pulsed-field gel electrophoresis (PFGE) and Southern hybridization for detection of genes encoding known virulence factors, ribosomal RNA gene spacer restriction fragment length polymorphism types (RSTs), ospC group determination, and sequencing of the variable dbpA and ospC genes. We show that two strains isolated from the same tick and both originally named N40 are in fact very distinct. Furthermore, we failed to detect bbk32, which encodes a fibronectinbinding adhesin, in one {"}N40{"} strain. Thus, two distinct strains that show different plasmid profiles, as determined by PFGE and PCR, were isolated from the same tick and vary in their ospC and dbpA sequences. However, both belong to group RST3B.",
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Detection of established virulence genes and plasmids to differentiate Borrelia burgdorferi strains. / Chan, Kamfai; Casjens, Sherwood; Parveen, Nikhat.

In: Infection and immunity, Vol. 80, No. 4, 01.04.2012, p. 1519-1529.

Research output: Contribution to journalArticle

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