Casting a shadow over dark field microscopy

Three years ago, I discussed a study by Laane and Mysterud, two researchers at the University of Oslo, who believed they had developed a reliable method for detecting Borrelia using dark field microscopy (1). This technique, in different variations, has been a popular subject for online videos and for Lyme diagnostics in the alternative sector (‘live blood analysis’). At the time, I already observed that it is not easy to determine whether structures that are visible under the microscope are actually Borrelia, or perhaps some other spirochete or even inanimate structures such as remnants of defective blood cells. The Norwegian researchers saw ‘spirochetes’ in the blood of chronic Lyme patients (who more often than not did not have Lyme according to official diagnostics), but they did not examine any healthy subjects; disregarding such a control group is a serious omission in scientific research. Some time later, the microscopy method was expanded to include detection of Babesia. Using their method, the researchers often came to very different conclusions than conventional diagnostics for Borrelia and Babesia.

The study caused considerable commotion in Norway and the researchers were even suspended from the university; presumably mainly because of the suggestion that conventional Lyme diagnostics based on serology is flawed. As a result of the controversy and pressure exerted by patient organizations, the microscopy method was further investigated by the Norwegian CDC, in cooperation with government agencies from several other EU countries and in consultation with the original researchers. For this follow-up, blood samples of both ‘chronic Lyme patients’ (most with a tick bite history, but without positive serology) and healthy subjects without a tick bite history were ‘blindly’ examined with the microscopy method, with conventional Lyme serology and with Borrelia PCR tests at five different laboratories. Each test lab also received a blood sample of a healthy subject to which Borrelia or Babesia had been added in a ratio of about 1 parasite per 20 red blood cells (a quantity that should be clearly visible under a microscope, but that is roughly a million times higher than usual with chronic Lyme).

With some delay, the results of this ‘counter-study’ have finally been published (2). These results show that the microscopy method is unsatisfactory for Lyme diagnostics: the researchers found ‘Borrelia spirochetes’ and ‘Babesia’ in most of the subjects, but more in healthy subjects than in chronic Lyme patients. This presumes that they do not perceive Borrelias, but dead structures or at most other (harmless) micro-organisms. No attempt was made to discover what the visible structures under the microscope actually were.

Nevertheless, this counter-study does present several problems. For instance, the results of the PCR tests of the different labs were very dissimilar and only the blood samples with added Borrelias were correctly diagnosed by all labs. Using PCR they found – just as with the microscopy method of Laane and Mysterud – more ‘positives’ in healthy subjects than in Lyme patients. Which of the PCR labs is correct and how to explain such large variations remains unclear. The authors suggest that PCR is simply not suitable for this type of diagnostics. The positive PCR results were not confirmed by conventional Lyme serology and this settles it for them: they were false-positive PCR results due to contamination or inadequately specific tests. Although the study superficially mentions which PCR technique was used by the various labs (all different methods, sometimes including sequencing confirmation), it does not mention which labs were concerned. The fact that a lab is capable of performing Borrelia PCR tests does not mean that they are good at it. A true assessment of the microscopy and PCR results is therefore impossible.

In the introduction to their article, the authors state that Lyme serology has a 70-90% sensitivity in the earliest spotlight2stages and a sensitivity of almost 100% in later stages, which is a belief that is shared by many Dutch medical microbiologists, but one that is absolutely incorrect and therefore very misleading; the sensitivity of serology (according to the standard two-step protocol) is much lower. The authors further state that disseminated Lyme and Babesia infections are extremely rare in Norway, but that there could be a high ‘background exposure’ to Borrelia of up to 20%. This percentage is likewise misleading, but the authors’ message is clear: a positive Lyme test has little meaning, ‘it happens all the time.’ Considering the numerous dubious propositions, you might wonder whether the selection of healthy subjects was perfectly legitimate, because this aspect is also impossible to verify. This creates the impression that the study was primarily designed to once again confirm the IDSA doctrine, to discredit microscopy and PCR diagnostics for Lyme and to emphasize that serology is the only acceptable diagnostic method.

The chief conclusion of this study seems to me that Borrelia PCR diagnostics urgently needs to be validated, for example through yearly proficiency testing with blood samples to which various Borrelia species have been added (in realistic concentrations, otherwise it is meaningless, and organized by an independent body). Even though such quality tests are relatively easy to arrange, they remain absent, similar to what is happening in the US. As a result, IDSA and its henchmen can continue to maintain that PCR is an unreliable method for Lyme diagnostics and patients are compelled to keep relying on outdated serology. In short, this publication of the Norwegian CDC feels more like shameless IDSA propaganda than a decent counter-study. 0-0 for science 🙁

  1. Borrelia detection using microscopy
  2. Validate or falsify: Lessons learned from a microscopy method claimed to be useful for detecting Borrelia and Babesia organisms in human blood

Published: February, 2016

Aangepast: 8 augustus 2020