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Detected Borrelia Burgdorferi in my Serum (Self Experiment) via PCR

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Lyme Disease
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Posted 6/9/2017 2:59 AM (GMT 0)
Hi all,

thought I should share this was you. Some of you helped me understand my immunoblott test since I wasn't really familiar with the diagnosis criteria and with my symptoms I am considered positive for lyme (Bands 23 and 41 IgG).
The lab I am with does not perform tests for lyme but we are involved in research. I went a head and did an experiment based on this study from American Journal of Clinical Pathology (Am J Clin Pathol. 2010 Apr;133(4):569-76. Increased sensitivity and specificity of Borrelia burgdorferi 16S ribosomal DNA detection.) on which they used 2 rounds of PCR with specific Borrelia Bur. DNA primers. Essentially it would amplify the ribosomal DNA from the lyme bacteria if you have it.

I drew blood, removed serum, and extracted potential DNA from serum using a very general method which would probably give low yield (i.e. it is not optimized to extract DNA from serum).
As you can see from the attached Image, the expected size band (~293 bp) is clear in the second round of PCR using Borrelia Bur specific DNA primers. Not sure how long the image hosting site will allow the image to stay, will re upload if it goes down as I couldn't do it directly through the form
[img]https://ibb.co/kp8Hha[\img]

Now this is interesting because I did not see any spirochete in the live blood of this same blood sample, but the bacteria is releasing DNA from somewhere. I wonder if I workout and get the usual herx if I would see more DNA amplified. I guess this is a direct proof of infection.
The good thing about doing your own experiment is that you can mess with the PCR amplification cycles compared to labs which would have a standard cycle that may give you false negative if your DNA count in the blood is very low. Also, this experiment used a different PCR type (Nested) and not the regular one round PCR.

-Immunity
Posted 6/9/2017 3:13 AM (GMT 0)
Very interesting Immunity!

-p
Posted 6/9/2017 9:17 AM (GMT 0)
You are lucky to have access to such a lab .maybe you can help others with testing someday. Where in the globe are you?
Posted 6/9/2017 12:43 PM (GMT 0)
I am thankful that I have that type of training and resources. I doubt I would've been positive through standard PCR with other labs unless the bacterial load in the blood was relatively high. This method that I used is about 100-1000 fold more sensitive than some regular PCR methods that detects lyme agents. We reside in Illinois, I would include more detail about me or the lab but don't since this is a public forum. I am happy to chat with anyone privately though smile. In the near future I am hoping to help develop more sensitive molecular tests for these types of infections.
Posted 6/9/2017 1:50 PM (GMT 0)
" I am happy to chat with anyone privately though smile. In the near future I am hoping to help develop more sensitive molecular tests for these types of infections."

This would be awesome, Immunity! I do hope that you get to fulfill your dream of being able to help make more sensitive testing!!! We so need that!

And, for those interested - here is a link to his photo: /ibb.co/kp8Hha
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