• Welcome to Phoenix Rising!

    Created in 2008, Phoenix Rising is the largest and oldest forum dedicated to furthering the understanding of, and finding treatments for, complex chronic illnesses such as chronic fatigue syndrome (ME/CFS), fibromyalgia, long COVID, postural orthostatic tachycardia syndrome (POTS), mast cell activation syndrome (MCAS), and allied diseases.

    To become a member, simply click the Register button at the top right.

Falsified: Microscopy method is not useful for detecting Borrelia & Babesia

deleder2k

deleder2k

Senior Member
Messages
1,129
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.
Click to expand...

http://www.tandfonline.com/doi/abs/10.3109/23744235.2016.1144931?journalCode=infd20
 
Asa

Asa

Senior Member
Messages
179
Devil's advocate: If not borrelia, what are the structures? And/or why are they appearing or seeming to appear? Does PCR always identify borrelia? If so, why isn't it used for patient diagosis/testing?

I'm just trying to get a very general (emphasis on general (!)) understanding, should anyone already know enough to provide such general/easy info. :)
 
A

alicec

Senior Member
Messages
1,572
Location
Australia
"PCR analysis detected Borrelia DNA in one sample of the patient group and in eight samples of the control group"

Doesn't sound like a very specific PCR technique - what an extraordinary false positive rate. Still if this technique couldn't find a positive in the LM-verified samples, nothing would!
 
deleder2k

deleder2k

Senior Member
Messages
1,129
"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."

The patients were previously interpreted by the LM method to have borrelia and or babesia. They was probably diagnosed by the LM method, or by shabby pseudo tests from Germany before entering. It doesn't sound strange that their results in both groups were low.
 
Antares in NYC

Antares in NYC

Senior Member
Messages
582
Location
USA
alicec said:
"PCR analysis detected Borrelia DNA in one sample of the patient group and in eight samples of the control group"

Doesn't sound like a very specific PCR technique - what an extraordinary false positive rate. Still if this technique couldn't find a positive in the LM-verified samples, nothing would!
Click to expand...
My thoughts exactly. Unless I'm missing something, this study is also exposing PCR as highly unreliable.
 
Dufresne

Dufresne

almost there...
Messages
1,039
Location
Laurentians, Quebec
Granted I'm no ace when it comes to parsing these things but it looks like their PCR is bogus. And that they actually imply the PCR results falsified the method. WTF? I would say finding 66% positive in the patient group and 85% in healthy controls virtually falsified the method. It seems to me both the methods were proven to be garbage. Correct me if I'm wrong.
 
A

alicec

Senior Member
Messages
1,572
Location
Australia
Antares in NYC said:
this study is also exposing PCR as highly unreliable.
Click to expand...

I was a bit surprised by the results because I did think PCR was very reliable. The main problem seems to be the low levels of bacteria in biological samples, the presence inhibitors and potential loss of bacteria from sample handling. These can all result in a potentially high number of false negatives.

I finally got around to doing a little research - not exhaustive, not enough energy. I found a very comprehensive review here It's a bit old but does look at PCR in different tissues/body fluids and gives an idea of sensitivity, false positives etc.

There's no need to wade through all the detail, you can just look at the tables.

The only test sample where there seems to be an issue with false positives is urine. They do comment in the text that there might be cross-reactivity with other infectious agents in urine, though why it is more of an issue in this medium is a bit of a mystery.

In any case, in the section on avoiding contamination, the paper emphasises what I think is really the problem with the study which is the subject of this thread. The amount of specific DNA in the test samples is very small so even a tiny amount of contamination of DNA from the testing lab could affect the outcome. The level of care needed to eliminate this type of contamination is common in research labs but might not be elsewhere.

I don't think this study is showing that PCR is necessarily unreliable, just that it can certainly be unreliable in the wrong hands.
 
Research 1st

Research 1st

Severe ME, POTS & MCAS.
Messages
768
deleder2k said:
"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."

The patients were previously interpreted by the LM method to have borrelia and or babesia. They was probably diagnosed by the LM method, or by shabby pseudo tests from Germany before entering. It doesn't sound strange that their results in both groups were low.
Click to expand...

Hello.

Please can you elucidate who exactly these 'shabby pseudo tests' from Germany are from please, as in the laboratory names?

I'd like to know the names of the labs you believe are ''shabby pseudo tests'', because so far, all the labs I've seen in Germany are of a reliable quality standard. (DAkkS accredited - German quality standard) and others are associated to German Universities.

Thanks.
 
deleder2k

deleder2k

Senior Member
Messages
1,129
I think this has been discussed on the forum previously. Many labs use unverified tests. Shabby was probably not a wise word to use. I meant undocumented/unverified.
 
bertiedog

bertiedog

Senior Member
Messages
1,738
Location
South East England, UK
I would like to add with regard to the German labs like Infectolab which is linked to BCA are in fact accredited and it was announced on Facebook via the UK Lyme group who has Jon Caldwell as a member that the NHS will now accept the results of positive tests for borrelia because they are satisfied with this lab at least.

Apparently Armin's lab has applied for the same accreditation and it is likely to be granted too so perhaps some people on here will have to review their attacks on these German labs at least.

Pam
 
B

barbc56

Senior Member
Messages
3,657
Accreditation does not mean a test is valid. This may be changing as there's a call for listing these tests as medical devices where accreditation is more strictly defined. This is in the US so don't know if US accreditation is comparable to other countries.

There is already some regulatory oversight of labs that offer LDTs. A federal law, The Clinical Laboratories Improvement Act (CLIA), requires assessment of whether a lab is able to test according to instructions and detect what is intended to be detected, that is, the test’s analytical validity. However, a key element is missing in that evaluation: clinicalvalidity. Does the test accurately diagnose or predict the risk of a particular clinical outcome
Click to expand...

https://www.sciencebasedmedicine.org/new-fda-regulatory-role-threatens-bogus-diagnostic-tests/
 
deleder2k

deleder2k

Senior Member
Messages
1,129
I spoke to a skeptical doctor about the low rate of sensitivity in PCR blood (3.4%) and this study. He said that if they could see lyme bacterias in a microscope, then the PCR test would sure as hell be positive.
 
D

duncan

Senior Member
Messages
2,240
Not sure if that logic holds up, @deleder2k . The efficacy of PCR was gauged at times against culture-positive cases, and I don't think it did all that well. I think its range was 10 to 60%?

Sorry, no studies to refer to at the moment to back my admittedly shaky memory.
 
You must log in or register to reply here.