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A simple method for the detection of live Borrelia spirochaetes in human blood 2013-08-12 (+)

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A simple method for the detection of live Borrelia spirochaetes in human blood using classical microscopy techniques
Morten M. Laane1, Ivar Mysterud2,*
1 Department of Molecular Biosciences, University of Oslo, P.O. Box 1041 Blindern, 0316 Oslo, Norway
2 Department of Biology, University of Oslo, P.O. Box 1066 Blindern, 0316 Oslo, Norway

We have developed a simple method for the detection of live spirochaete stages in blood of patients where chronic borreliosis is suspected. Classic techniques involving phase-contrast and fluorescence microscopy are used. The method is also quite sensitive for detecting other bacteria, protists, fungi and other organisms present in blood samples. It is also useful for monitoring the effects of various antibiotics during treatment. We also present a simple hypothesis for explaining the confusion generated through the interpretation of possible stages of Borrelia seen in human blood. We hypothesize that these various stages in the blood stream are derived from secondarily infected tissues and biofilms in the body with low oxygen concentrations. Motile stages transform rapidly into cysts or sometimes penetrate other blood cells including red blood cells (RBCs). The latter are ideal hiding places for less motile stages that take advantage of the host’s RBCs blebbing-system. Less motile, morphologically different stages may be passively ejected in the blood plasma from the blebbing RBCs, more or less coated with the host’s membrane proteins which prevent detection by immunological methods.

Since the discovery of Borrelia burgdorferi, the Lyme disease spirochaete, borreliology has evolved from a microbiological curiosity to a major branch of bacterial pathogenesis research. Lyme disease is a global health problem; indeed few infectious diseases have garnered more sustained attention from the scientific and, notably, the lay media [1]. Borrelia-related diseases are a major challenge in medical research in many countries of the world, and we refer to comprehensive sources for the history and characteristics of this dangerous disease [2]. The disease is of great relevance today, and will be even more important tomorrow [3].
The most serious challenges are connected to diagnosis. The relatively large Borrelia, i.e. B. burgdorferi, is in general not readily detectable in blood smears of varying thickness from Lyme disease patients and suspected host animals. Yet blood engorgement from infected host result in up to 100% infected ticks [4].

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