Bacterial Infections and CSF Research

If it's on your mind and it has to do with multiple sclerosis in any way, post it here.

Bacterial Infections and CSF Research

Postby Shayk » Mon Oct 02, 2006 6:51 pm

Hi all

Given all the interest in infections here's another study from Ectrims 2006 that I'm also surprised no one has posted. :roll:
PCR for bacteria in multiple sclerosis cerebrospinal fluid
J.W. Lindsey, S. Patel (Houston, USA)

Objective: The objective of this study was to use a sensitive PCR method to test for the presence of seven different groups of bacteria in the cerebrospinal fluid (CSF) of multiple sclerosis patients.

Background: The etiology of multiple sclerosis is currently unknown, but many features of the disease suggest an infectious cause. The clinical course, the inflammatory infiltrates seen on histology, and the oligoclonal bands in the cerebrospinal fluid are all consistent with a recurrent or persistent infection. Similar diseases, such as acute transverse myelitis, acute disseminated encephalomyelitis, and Guillain-Barre syndrome, are linked to infections. And chronic infection with Treponema or Borrelia can cause neurologic disease.

Methods: We designed nested sets of PCR primers specific for the 16S ribosomal DNA of spirochetes, campylobacter, mycoplasma, chlamydia, bartonella, mycobacteria, and streptococcus. Each set of primers is designed to amplify DNA from all members of the desired group without amplifying DNA from common laboratory contaminants. We extracted DNA from the CSF of 10 patients with relapsing-remitting MS, 10 patients with primary progressive MS, and 9 controls. We amplified the DNA with nested PCR, and visualized the PCR products on agarose gels. Each experiment included both MS patients, controls, and a saline negative control. We defined the sensitivity of our method using serial dilutions of known amounts of bacterial DNA.

Results: The majority of the CSF specimens were negative for bacteria with all sets of primers. Three MS specimens and one control had a faint band of the correct size with the spirochete primers. Sequencing identified this PCR product as coming from a propionibacterium rather than a spirochete. Further experiments with primers specific for that sequence demonstrated that it was present at low concentrations in all specimens and is likely a laboratory contaminant. The sensitivity of the method was excellent. We could detect 10 copies or less of bacterial DNA per PCR reaction. This corresponds to a concentration of 200 bacteria per ml of CSF.
Conclusions: Using a very sensitive PCR method, we were unable to find evidence for the presence of any of the seven tested groups of bacteria in the CSF of MS patients.
This study supported in part by the National Multiple Sclerosis Society

Now, I thoroughly expect some will readily dismiss the study, particularly as it might pertain to the controversies surrounding cpn, difficulties culturing it, and the discrepancies among labs. I do know (surprise) that some labs seem to find cpn while others don't. This one apparently didn't find evidence of cpn or of the other bacteria that were tested.

I'm thankful that there's continued research in this area. And, the beat goes on......as does the controversy I'm sure. :wink:

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Postby mrhodes40 » Mon Oct 02, 2006 8:15 pm

Yes we need more research in this area. Importantly the page said this
Background: The etiology of multiple sclerosis is currently unknown, but many features of the disease suggest an infectious cause. The clinical course, the inflammatory infiltrates seen on histology, and the oligoclonal bands in the cerebrospinal fluid are all consistent with a recurrent or persistent infection. Similar diseases, such as acute transverse myelitis, acute disseminated encephalomyelitis, and Guillain-Barre syndrome, are linked to infections. And chronic infection with Treponema or Borrelia can cause neurologic disease


The fact that they did not find much may not mean as much as we wish it did though
The most frequent and serious manifestation of disseminated Lyme borreliosis is neuroborreliosis. PCR was applied to 190 patients with untreated and confirmed neuroborreliosis. B. burgdorferi DNA was detectable in 17-21% of CSF samples from patients with neuroborreliosis. In patients with very early neuroborreliosis (< 2 weeks), still being negative for specific intrathecal antibody synthesis, a positive PCR was more frequent than in patients with longer disease duration.


interesting that the longer duration resulted in less PCR positivity- it is odd. Certainly CPn in MS would be a long term infection. But take note this paper was confirmed disease with other manifestations and we have a really low positivity in the CSF. I don't know about you but 17% seems flat awful to me considering these patients are really ill with an already identified serious illness can you imagine if this was the only way they would know they had this?? 83% would be told Nope, negative. Are you kidding me?

This page http://tinyurl.com/nbwcy mentions the problem of culture speifically
no agent has yet emerged with any consensus as the cause of MS. This controversy is due to a number of factors, including lack of specificity of an agent to MS, lack of reproducibility in other laboratories, inappropriate controls, laboratory contamination and lack of a standard and easily reproducible assay system.


Another recent paper
Chlamydia pneumoniae infection associated with enhanced MRI spinal lesions in multiple sclerosis.Hao Q, Miyashita N, Matsui M, Wang HY, Matsushima T, Saida T.
Department of Neurology, Center for Neurological Diseases, Utano National Hospital, Kyoto, Japan.

Cerebrospinal fluid (CSF) from 66 patients with multiple sclerosis (MS) and 25 patients with other neurological diseases (OND) were examined for the infection of Chlamydia pneumoniae by culture, polymerase chain reaction (PCR) assay, and determination of antibodies to C. pneumoniae. PCR was positive not only in 9 of 28 (32%) patients with MS but also in 2 patents with inflammatory disorders in 15 (13%) OND controls (p = 0.18). Viable C. pneumoniae was isolated from one patient with MS and one with paraneoplastic encephalomyelitis. C. pneumoniae could be detected only in cell-containing CSF. In MS, enhanced spinal magnetic resonance imaging (MRI) lesions were detected in all of four PCR-positive patents but none of five PCR-negative patients, and the difference was significant (p = 0.0079). However, no correlation was found between enhanced brain MRI lesions and CSF C. pneumoniae DNA. Elevated titers of anti-C. pneumoniae IgG were detected in CSF in 13 of 66 (20%) patients with MS and 1 of 25 (4%) OND controls (p = 0.064). CNS C. pneumoniae infection is not uncommon in MS as well as in other inflammatory disorders of the nervous system. The association of active spinal lesions with Chlamydia in CSF collected by lumber puncture suggests the detection of a recent infection. On the other hand, the lack of association of active MS brain lesions with CSF Chlamydia and the presence of PCR-positive patents who are clinically stable and have no enhancing MRI lesions imply the existence of a chronic infectious process.

PMID: 12356213 [PubMed - indexed for MEDLINE]


So of course we are back to the old you have your scientist and I have a different one, so who's got the right guy? This hasn't been resolved and we keep seeing papers on this as there IS a lot of material on both sides. This is science in progress. I read a paper recently that stated that it takes a 33% agreement with a new paradigm before the "tipping point" occurs and the old thought gives way to the new idea We have a ways to go before we reach consensus :wink:
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Postby mrhodes40 » Mon Oct 02, 2006 9:00 pm

I forgot to share one my favorite reads on this subject
Hammerschlag MR.
Division of Pediatric Infectious Diseases, SUNY Downstate Medical Center, Brooklyn, NY 11203-2098, USA. mhammerschlag@pol.net

One of the major characteristics of Chlamydia spp. is its ability to cause prolonged, often subclinical infections. Chronic, persistent infection with Chlamydia pneumoniae has been implicated in the pathogenesis of several chronic diseases initially not thought to be infectious, including asthma, arthritis and atherosclerosis. C. pneumoniae is susceptible in vitro to a wide range of antimicrobial agents that target either protein or DNA synthesis, including macrolides, ketolides, tetracyclines, quinolones and rifamycins. Practically all treatment studies evaluating presented or published to date have used serology alone for diagnosis of C. pneumoniae infection, which only provides a clinical end point. The results of several treatment studies that did perform culture found that erythromycin, azithromycin (Zithromax, clarithromycin (Biaxin, levofloxacin (Levaquin and moxifloxacin (Avelox had a 70 to 90% efficacy in eradicating C. pneumoniae from the respiratory tract of children and adults with pneumonia. Persistence of the organism does not appear to be due to the development of antibiotic resistance. However, one cannot extrapolate from this experience to the treatment of chronic C. pneumoniae infection, especially cardiovascular disease. As there are no reliable serologic markers for chronic or persistent C. pneumoniae infection, it cannot be determined who is infected and who is not, which means that it cannot be assumed that any effect seen is due to successful treatment or eradication of C. pneumoniae.

PMID: 15482145 [PubMed - indexed for MEDLINE]

Essentially this author who wrote this expert review is saying we can't culture persistent infection. Her point is that since we can't culture it we can't know if treated people are better because of the treatment. But note she lists several 'thought to be' autoimmune diseases as probably CPn diseases. Also note she is the one who wrote some years before this paper, she wrote this negative article on the idea of MS being CPn http://cdli.asm.org/cgi/content/full/10 ... d=12965938. Her later assertion that I quoted in complete above makes one wonder if she is still so sure MS cannot be CPn since she is saying now that persistent infection is probably causing other chronic diseases like arthritis and asthma.
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