Microscopy Method For Detecting Lyme Under Fire.


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

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 beBorrelia- 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.

Only one problem...PCR isn't a reliable way to find Lyme disease either.

From Medscape:

This leads to the question. is PCR useful for the diagnosis of Lyme disease? In general, the answer is no.

Borrelia burgdorferi, the spirochete that causes Lyme disease, concentrates in collagen-rich connective tissues. Although spirochetes initially disseminate from the site of an infected tick bite via the blood, the bloodborne phase is relatively brief and the concentration of spirochetes is quite low. In fact, PCR detectsBorrelia DNA in the blood of fewer than half of patients in the early acute stage of disease when the erythema migrans rash is present. By the time symptoms of Lyme disease have been present for a month or more, spirochetes can no longer be found in blood. Similarly, PCR testing of cerebral spinal fluid (CSF) specimens is not clinically useful. PCR testing of CSF is positive in only about one third of US patients with early neuroborreliosis, and it is even less sensitive in patients with late neurologic disease. Urine is not a suitable sample for PCR testing at any stage of Lyme disease.

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