Validate or falsify: Lessons learned from a microscopy method claimed to be useful for detecting Borrelia and Babesia organisms in human blood
Language English Country Great Britain, England Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- Keywords
- Babesia spp., Borrelia burgdorferi sensu lato, Lyme borreliosis, Lyme disease, PCR, babesiosis, microscopy,
- MeSH
- Babesia genetics isolation & purification MeSH
- Babesiosis blood diagnosis parasitology MeSH
- Borrelia genetics isolation & purification MeSH
- Child MeSH
- DNA, Bacterial analysis MeSH
- Adult MeSH
- Infant MeSH
- Middle Aged MeSH
- Humans MeSH
- Lyme Disease blood diagnosis microbiology MeSH
- Microscopy methods standards MeSH
- Adolescent MeSH
- Young Adult MeSH
- Polymerase Chain Reaction MeSH
- Child, Preschool MeSH
- DNA, Protozoan analysis MeSH
- Aged MeSH
- Sensitivity and Specificity MeSH
- Animals MeSH
- Check Tag
- Child MeSH
- Adult MeSH
- Infant MeSH
- Middle Aged MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Child, Preschool MeSH
- Aged MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- DNA, Bacterial MeSH
- DNA, Protozoan MeSH
BACKGROUND: A modified microscopy protocol (the LM-method) was used to demonstrate what was interpreted as Borrelia spirochetes and later also Babesia sp., in peripheral blood from patients. The method gained much publicity, but was not validated prior to publication, which became the purpose of this study using appropriate scientific methodology, including a control group. METHODS: Blood from 21 patients previously interpreted as positive for Borrelia and/or Babesia infection by the LM-method and 41 healthy controls without known history of tick bite were collected, blinded and analysed for these pathogens by microscopy in two laboratories by the LM-method and conventional method, respectively, by PCR methods in five laboratories and by serology in one laboratory. RESULTS: Microscopy by the LM-method identified structures claimed to be Borrelia- and/or Babesia in 66% of the blood samples of the patient group and in 85% in the healthy control group. Microscopy by the conventional method for Babesia only did not identify Babesia in any samples. PCR analysis detected Borrelia DNA in one sample of the patient group and in eight samples of the control group; whereas Babesia DNA was not detected in any of the blood samples using molecular methods. CONCLUSIONS: The structures interpreted as Borrelia and Babesia by the LM-method could not be verified by PCR. The method was, thus, falsified. This study underlines the importance of doing proper test validation before new or modified assays are introduced.
c Section for Virology Norwegian Veterinary Institute Oslo Norway ;
Department of Bacteriology and Immunology Norwegian Institute of Public Health Oslo Norway ;
Department of Medical Microbiology Sørlandet Hospital Health Enterprise Kristiansand Norway ;
f Department of Engineering and Science University of Agder Kristiansand Norway ;
g Research Unit Sørlandet Hospital Health Enterprise Kristiansand Norway ;
References provided by Crossref.org
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