ArticleEuropean journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology1996
Single-tube nested polymerase chain reaction assay based on Flagellin gene sequences for detection of Borrelia burgdorferi sensu lato.
Article in European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology, 1996. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- A two year prospective study to compare culture and polymerase chain reaction amplification for the detection and diagnosis of Lyme borreliosis.Molecular pathology : MP · 1997Trial
- Molecular survey ofCurrent research in parasitology & vector-borne diseases · 2026Article
- Transovarial and transstadial transmission of Borrelia valaisiana, the pathogen of Lyme borreliosis, in the sheep tick, Ixodes ricinus L.Experimental & applied acarology · 2024Article
- Article
- Detecting the Lyme Disease Spirochete, Borrelia Burgdorferi, in Ticks Using Nested PCR.Journal of visualized experiments : JoVE · 2018Article
- Occurrence ofJournal of the South African Veterinary Association · 2017Article
- Comparison of MKP and BSK-H media for the cultivation and isolation of Borrelia burgdorferi sensu lato.PloS one · 2017Article
- Molecular detection of zoonotic tick-borne pathogens from ticks collected from ruminants in four South African provinces.The Journal of veterinary medical science · 2016Article
- Pathogens vectored by the tick, Dermacentor reticulatus, in endemic regions and zones of expansion in Poland.Parasites & vectors · 2015Article
- Agents of human anaplasmosis and Lyme disease at Camp Ripley, Minnesota.Vector borne and zoonotic diseases (Larchmont, N.Y.) · 2011Article
- Article
- Single-tube nested PCR using immobilized internal primers for the identification of dengue virus serotypes.Journal of virological methods · 2007Article
- Use of a suspension array for rapid identification of the varieties and genotypes of the Cryptococcus neoformans species complex.Journal of clinical microbiology · 2005Article
- Occurrence of genes associated with enterotoxigenic and enterohemorrhagic Escherichia coli in agricultural waste lagoons.Applied and environmental microbiology · 2004Article
- Direct molecular typing of Borrelia burgdorferi sensu lato species in synovial samples from patients with lyme arthritis.Journal of clinical microbiology · 2000Article
- Human disease in Europe caused by a granulocytic Ehrlichia species.Journal of clinical microbiology · 1997Article
- PCR in laboratory diagnosis of human Borrelia burgdorferi infections.Clinical microbiology reviews · 1997Review
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Authors and funding
5 authors.
Funding
Abstract
An inherent drawback of nested PCR systems to increase sensitivity of PCR-based assays is that tubes must be opened after the first round of amplification in order to transfer template molecules to the second amplification reaction; this procedure introduces the risk of carry-over contamination of negative specimens. To obviate this disadvantage, a nested PCR assay for detection of Borrelia burgdorferi in which both amplifications are performed in a single tube that remains closed throughout the entire process was devised. The assay is based on flagellin gene sequences with previously determined species-wide and species-specific properties. The nested PCR system proved to be 1000 times more sensitive than the conventional assay. Using the nested PCR system, ten spirochaetes could be routinely detected by agarose gel electrophoresis alone, whereas the conventional PCR system could detect only 10(4) spirochaetes under these conditions. After Southern transfer on amplification products and hybridization with 320- or chemiluminescent-labeled probes, the nested PCR system could easily detect a single spirochaete by both means, whereas the sensitivity of the conventional PCR assay varied from 10(1) (32P) to 10(3) (chemiluminescence) spirochaetes. This single-tube nested PCR system should be a useful addition to the current range of diagnostic assays for Lyme borreliosis.
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