Eva Sapi study about new Borrelia blood culture test

Topics with information and discussion about published studies related to Lyme disease and other tick-borne diseases.
X-member
Posts: 6888
Joined: Mon 30 Jul 2007 18:18

Re: Eva Sapi study about new Borrelia blood culture test

Post by X-member » Sun 9 Mar 2014 2:21

nnecker, maybe it is easier to culture the bacteria if it done on blood from the right patients? (Not only after a very early treatment?)
Last edited by X-member on Sun 9 Mar 2014 2:57, edited 1 time in total.

User avatar
LHCTom
Posts: 341
Joined: Mon 22 Oct 2012 4:18

Re: Eva Sapi study about new Borrelia blood culture test

Post by LHCTom » Sun 9 Mar 2014 2:48

You seem to be a very smart guy.I am just a lay person.Can you explain why it is so easy for Sapi to culture viable spirokeets after abx treatment and impossible for everyone else,and they even have the advantage of using tissue?
They don't easily cultivate spirochetes from the patients. I suspect they have a quality control problem. I believe they have a contamination problem but I remain hopeful. I believe the Johnson paper explained the results in this study rather well. I suspect once they clean up the contamination, they will also find it difficult to culture post treatment. I tend to think Bathold is on the right track. I supect in most cases spirochetes that survive treatment and a full immune response, downregulate and become uncultivable and go into hiding not unlike the herpes virus strategy. I use the words "believe" and "suspect" because much of the evidence is inconsistent at best and still incomplete.
The greater the ignorance, the greater the dogmatism.

Attributed to William Osler, 1902

X-member
Posts: 6888
Joined: Mon 30 Jul 2007 18:18

Re: Eva Sapi study about new Borrelia blood culture test

Post by X-member » Sun 9 Mar 2014 8:36

nnecker wrote earlier:
Can you explain why it is so easy for Sapi to culture viable spirokeets after abx treatment and impossible for everyone else, and they even have the advantage of using tissue?
Borrelia burgdorferi detected by culture and PCR in clinical relapse of disseminated Lyme borreliosis.

http://www.ncbi.nlm.nih.gov/pubmed/1044 ... t=Abstract

A quote:
PCR was positive from the plasma of 14 patients 0-30 months after discontinuation of the treatment, and 12 of these patients had a clinical relapse. In addition, Borrelia burgdorferi was cultured from the blood of three patients during the follow-up. All three patients belonged to the group with relapse, and two of them were also PCR positive.

nnecker
Posts: 215
Joined: Wed 19 Dec 2012 22:57

Re: Eva Sapi study about new Borrelia blood culture test

Post by nnecker » Sun 9 Mar 2014 13:50

Tom,

This is the way I see it.Feel free to make any corrections if I am wrong about something.Lets first look at a mouses ear after 22 days of infection in this study.(photo a)

http://www.jci.org/articles/view/58813/figure/1

That seems like a pretty intense infection of spirokeets.Barthold let this go on for four months.It just seems to me that spirokeets would be oozing out of the mouses pours by then.lol.It also seems to me it's not where the spirokeets are hiding.Where could they not be hiding with this being such an intense infection?

Yet after 25 days of abx treatment and 12 months of tissue culturing,Barthold can't find one f-ing viable spirokeet?

Now when you look at photo b,look how many spirokeets die just after ONE DAY of abx use.And of course in this study they did not find sh-t either.Now in your case,you were taking all these abx's and were still on them when you gave Sapi blood,and then she magically made viable spirokeets appear from everywhere.

Why are these two scenarios so different.How did this happen?Is it because lab animals and humans are so much different?If this is the case.Are there any other examples of this happening with other pathogenic bacteria between lab animals and humans?

You talk about the herpes virus strategy,is this the new fad now?Is the cyst, biofilm fad now finally over?After you took all your abx's do you now believe you still have viable spirokeets in your body?And as far as the herpes strategy goes,does this mean at some point viable spirokeets will reappear in your body?
Last edited by nnecker on Sun 9 Mar 2014 17:01, edited 1 time in total.

Margherita
Posts: 192
Joined: Thu 27 Sep 2012 18:22

Re: Eva Sapi study about new Borrelia blood culture test

Post by Margherita » Sun 9 Mar 2014 16:27

Dear LHCTom,

I'm very interested in the new DNA Lymetest you referred to. But what about patients with persisting Lyme symptoms? Does this test distinguish recent DNA from old DNA (as you explained in other posts)?
And what about the acceptance of this test in Europe? Can you tell me more about it (maybe by dedicating a new, special topic to it in order not to interfere with this Sapi matter)?

Thanks in advance and have a nice day.

X-member
Posts: 6888
Joined: Mon 30 Jul 2007 18:18

Re: Eva Sapi study about new Borrelia blood culture test

Post by X-member » Sun 9 Mar 2014 17:37

nnecker wrote:
You talk about the herpes virus strategy,is this the new fad now?Is the cyst, biofilm fad now finally over?After you took all your abx's do you now believe you still have viable spirokeets in your body?And as far as the herpes strategy goes,does this mean at some point viable spirokeets will reappear in your body?
You seem to be very sure that abx always works darn good (no matter how long the patient have had the infection), don't you?

User avatar
LHCTom
Posts: 341
Joined: Mon 22 Oct 2012 4:18

Re: Eva Sapi study about new Borrelia blood culture test

Post by LHCTom » Mon 10 Mar 2014 2:01

Hey nnecker... lighten up, we both have the distinction of being "trolls", flamed by wackjob "Elena Cook" and accused of being Edward McSweegan... dosen't that make us brothers in arms? I've seen your other posts and know you know how to spell spirokeets and are smarter than you pretend to be.


Cazy Elena - http://www.mdjunction.com/forums/lyme-d ... k-activist
This is the way I see it.Feel free to make any corrections if I am wrong about something.Lets first look at a mouses ear after 22 days of infection in this study.(photo a)
1) Interesting study but duel-photon imaging has limited depth of penentration and cannot see into all potential immune priviledged places.

2) It had an 8% culture detection rate in heavily treated mice - That's rather large given the extensive treatment.

3) It doesn't rule all potential atennuated uncultivable spirochetes beyond the duel-photon reach and un-tested areas.

4) Stopped the study way too soon. Should have let some of mice live a whole year and repeat the testing...
That seems like a pretty intense infection of spirokeets.Barthold let this go on for four months.It just seems to me that spirokeets would be oozing out of the mouses pours by then.lol.It also seems to me it's not where the spirokeets are hiding.Where could they not be hiding with this being such an intense infection?
Oozing out of poors... How descriptive... Would make a good cartoon!

There are lots of immune priviledged places out of reach of antibiotics, the duel-photon microscopy and not tested at post-mortum.
Yet after 25 days of abx treatment and 12 months of tissue culturing,Barthold can't find one f-ing viable spirokeet?
Now watch your language.... Elena is watching!

Not quite accurate. Read more carefully.

http://www.ncbi.nlm.nih.gov/pmc/article ... 086907.pdf
Despite the continued non-cultivable state, RNA transcription of multiple B. burgdorferi genes was detected in host tissues, flaB DNA was acquired by xenodiagnostic ticks, and spirochetal forms could be visualized within ticks and mouse tissues by immunofluorescence and immunohistochemistry, respectively. A number of host cytokines were up- or down-regulated in tissues of both saline- and antibiotic-treated mice in the absence of histopathology, indicating host response to the presence of non-cultivable, despite the lack of inflammation in tissues.
Now when you look at photo b,look how many spirokeets die just after ONE DAY of abx use.And of course in this study they did not find sh-t either.Now in your case,you were taking all these abx's and were still on them when you gave Sapi blood,and then she magically made viable spirokeets appear from everywhere.
I think you misunderstand my view of the Sapi/ALS culture. I don't believe its any better than done before and hiding attenuated non-cultivable spirochetes can't be cultured. But they can annoy the immune system causing a multitude of inflammatory driven sysmptoms.

It does show how effective the antibiotics are that reach the spirochetes.

You talk about the herpes virus strategy,is this the new fad now?Is the cyst, biofilm fad now finally over?After you took all your abx's do you now believe you still have viable spirokeets in your body?And as far as the herpes strategy goes,does this mean at some point viable spirokeets will reappear in your body?
I'm not talking about "the cyst, biofilm fad". The reason the Herpes virus cannot be killed by antivirals or the immune system is the HSV virus persists by becoming latent and hiding from the immune system in the cell bodies of neurons which antivirals nor the immune system can reach. There are many such hiding places in the body that would be difficult to detect.
The greater the ignorance, the greater the dogmatism.

Attributed to William Osler, 1902

X-member
Posts: 6888
Joined: Mon 30 Jul 2007 18:18

Re: Eva Sapi study about new Borrelia blood culture test

Post by X-member » Mon 10 Mar 2014 7:06

Maybe this information belong in this thread?

The Complexities of Lyme Disease

http://www.lymeneteurope.org/info/the-c ... me-disease
The worst problem for chronic Lyme patients is that, after they are treated with antibiotics, they are told they are cured even if they have a recurrence of symptoms. There is a persistent dogma in medicine that 28 days of IV antibiotics cures all Lyme Disease. In fact, the ongoing six-year-old Nantucket Island Lyme Treatment Study showed IV antibiotics to have the highest relapse rate in late Lyme disease! This was because doctors put too much faith in IV antibiotics as being so powerful, that they did not follow up IV's with oral antibiotics. The key to treating late Lyme appears to be the length of antibiotic treatment, not the method. If IV's are followed up by six months or more of oral antibiotics, the relapse rate dropped to 13%. (Dr. Leslie Fein MD, MPH, Magnarelli MD, MPH 96 LDF Conference)

Martian
Posts: 1944
Joined: Thu 26 Jul 2007 18:29
Location: Friesland, the Netherlands

Re: Eva Sapi study about new Borrelia blood culture test

Post by Martian » Mon 10 Mar 2014 17:55

The study by Sapi et al. on the new culture method was assessed by Johnson et al. from the CDC:
J Clin Microbiol. 2014 Mar;52(3):721-4. doi: 10.1128/JCM.01674-13. Epub 2013 Aug 14.

Assessment of new culture method for detection of borrelia species from serum of lyme disease patients.

Johnson BJ [1], Pilgard MA, Russell TM.

1. Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado, USA.

Abstract

A novel method of culturing spirochetes from the serum of U.S. Lyme disease patients was recently reported by Sapi and colleagues to have 94% sensitivity and 100% specificity for Borrelia species as assessed by microscopy and DNA sequence analysis of the pyrG gene (E. Sapi, N. Pabbati, A. Datar, E. M. Davies, A. Rattelle, and B. A. Kuo, Int. J. Med. Sci. 10:362-376, 2013).

The majority of the spirochetes described were related by pyrG sequences to species of Borrelia previously undetected in North American patients without a reported history of travel to Europe or Asia.

To better understand these unexpected findings, we determined pyrG sequences of the laboratory reference strains used by the investigators for method development and testing of culture medium.

Eighty percent (41/51) of the reported patient-derived pyrG sequences were identical to one of the laboratory strains, and an additional 12% (6/51) differed by only a single nucleotide across a 603-bp region of the pyrG gene.

Thus, false positivity due to laboratory contamination of patient samples cannot be ruled out, and further validation of the proposed novel culture method is required.

PMID: 23946519 [PubMed - in process]
The full text can be found here: http://jcm.asm.org/content/early/2013/0 ... M.01674-13

User avatar
LHCTom
Posts: 341
Joined: Mon 22 Oct 2012 4:18

Re: Eva Sapi study about new Borrelia blood culture test

Post by LHCTom » Mon 10 Mar 2014 21:37

I'm very interested in the new DNA Lymetest you referred to. But what about patients with persisting Lyme symptoms? Does this test distinguish recent DNA from old DNA (as you explained in other posts)?
And what about the acceptance of this test in Europe? Can you tell me more about it (maybe by dedicating a new, special topic to it in order not to interfere with this Sapi matter)?
I'll start a new topic with a little more detail... but it cannot distinguish DNA junk or fragment that contains a Borrelia 16S ribosomal RNA gene that's a year old or a new one from living Borrelia. It is nice because the sequencing almost guarantees its Borrelia and identifies the genotype even if its the relapsing fever B. miyamotoi commonly found but not detected by the antibody testing. It will only detect Borrelia if there are a large number of spirochetes in your blood or very lucky. That means its probably only useful to confirm Lyme pre-treatment and during the early phase. It would be interesting if they would accept semen samples so we could show the STD theory is bogus.

The use of the 16S ribosomal RNA gene is standard in bacterial identification. I suspect it would be accepted in Europe since the 16S ribosomal RNA gene is commonly used, the low temperature nested PCR is widely used, its been validated many times and there are 2 papers published on the specific implementation. The lab itself can always be suspect for contamination or quality control.


That being said, if Borrelia does hide and re-emerge and one is having a strong flare of symptoms, there is a small chance it could detect Lyme if the strong flare of symptoms goes along with a resurgence in spirochetes in the blood. It is a direct antigen detection method so does not suffer from the many antibody based test weaknesses but does have its own. At $150 for 3 PCR runs, its certainly worthy of trying as a confirmation early in an infection or late during extreme symptom flares while off antibiotics for a long time. Its the first time a research quality low temperature nested PCR with sequencing has been made available to clinicians. I would like to see as many chronic Lyme suffers as possible give it a try but only while off antibiotics and during a severe symptom flare.

They emailed me my results from 6 PCR runs and it was negative as expected. I was not having a svere symptom flare nor had I been antibiotics for a long time. I did have 6 Western Bot antibodies detected plus C6 peptide ELISA positive twice by 2 labs both using the Immunetics kit. So that's one data point.

more later
The greater the ignorance, the greater the dogmatism.

Attributed to William Osler, 1902

Post Reply