Muliple-Strain Bb Infections: Implications

Medical topics with questions, information and discussion related to Lyme disease and other tick-borne diseases.
duncan
Posts: 1370
Joined: Wed 5 Sep 2012 18:48

Muliple-Strain Bb Infections: Implications

Post by duncan » Tue 13 Aug 2013 10:20

I do not know if when I submit to an ELISA or Western Blot Lyme test, and if I draw a positive (or a negative for that matter), what strain of Borrelia that metric reports on. If I am in the U.S., I at least am pretty clear on the species, but the strain - not so much.

As I thought about this, questions came to mind:

Do all Labs test for the same strain, and only that strain? For instance, if I am tested through Labcorp, and that company employs a kit that reports on B31 (I have been told, but not verified), would that hold true for other labs? I believe the answer is no, or at least, not necessarily. Igenex leaps to mind as in I believe it tests for two strains. If some labs select one strain over another to test for, why? If I test positive for one variation, is that as good, in effect, as testing for any other strain, i.e., is it immaterial diagnostically which strain one contracts and is tested for?

Can you get labs to break out the specific strains one has? I have had many tests for Lyme. The vast do not provide data that specific, i.e., which strain(s) you are infected with. That being said, I also have tests which DO seem to specify the strain. Some seem to indicate multiple strains, and even break out each strain's specific bands when they involve WB's. Maybe I am misreading them, but I don't believe so.

Does it even matter? What are the ramifications of being infected with more than one strain? The first paper I found that even addresses the possibility of multiple strain infections was from 1999 and included Dattwyler. Sorry, no link. But, would being infected by multiple strains make it more difficult for one's body to clear the infections? Does treatment get a little trickier potentially? Or, will a conventional dose of abx take care of the whole bunch in a single collective swoop? Is there even a protocol for multiple strain infections? Or, to get darker and more speculative, if left untreated, could each strain develop unto its own peculiarities, and engender symptomalogies that are strain-specific, and become individually entrenched, and, well, you get the idea. It's the what-happens-if-you-let-one-strain-go-untreated scenario, potentially, times two. Or three.

Anyways. Some Lyme musings I figured I'd burden y'all with, to run with or ignore.

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

Re: Muliple-Strain Bb Infections: Implications

Post by LHCTom » Tue 13 Aug 2013 21:36

Do all Labs test for the same strain, and only that strain? For instance, if I am tested through Labcorp, and that company employs a kit that reports on B31 (I have been told, but not verified), would that hold true for other labs? I believe the answer is no, or at least, not necessarily. Igenex leaps to mind as in I believe it tests for two strains. If some labs select one strain over another to test for, why? If I test positive for one variation, is that as good, in effect, as testing for any other strain, i.e., is it immaterial diagnostically which strain one contracts and is tested for?
There are no commercial tests for Bb strain and it probably does not matter. There is little to none known about the relationship between Bb strain and the virulence, treatment and course of the infection. If a lab wants to get FDA approval for their Lyme tests, then they are required to use the B31 strain antigens. Labcorp almost certainly uses a kit based on the B31 strain. If you test positive then the strain is irrelevant. The issue is that the antigens used in the test lead to the test being sensitive to strain. If you are infected with a strain where its epitopes are sufficiently different from B31, then the test may fail. Now you have an uphill battle getting diagnosed if the test failed due to strain. You are now negative forever. This is one of the reasons the CDC 2T test is criticized. Igenex and Stony Brook use two strains in an attempt to reduce this strain sensitivity. Researchers including Wormser routinely mention genotype/strain sensitivity but to my knowledge, no studies have been done to identify actual human infections and the strain and then tested the CDC 2T test for sensitivity to real world varied strains. The C6 peptide test ( available at Igenex, Stony Brook and Quest) was designed in part to minimize strain sensitivity. So you can get the C6 peptide test here in the US as an adjunct to the CDC 2T test to try and catch strains that are missed. It too fails on some strains as reported by Barthold.

It seems very unlikely one person would have 2 strains but its certainly possible and has happened. It probably makes little difference to symptoms or treatment but its just not known. There is virulence variations by OspC type which varies by strain but its not a one to one relationship. So until a test arrives that provides strain or OspC type ( Barbour is working on one that might), little will remain known about how it affects the course of the disease in the real world. The European's have at least 3 different species which do tend toward different dominant symptoms. The US CC 2T test fails on their non-Bb species. The US has at least 3 other pathogenic species besides Bb. The Western blot does not provide strain information. The bands indicate the outer surface proteins or antigens in Borrelia found by the test. The only way to determine Bb strain is to test it with a multi-locus PCR. This is only done in research labs today. If you get the ALS culture, they will send the culture out to Eurofins for partial DNA sequencing. This however does not sequence enough to determine actual strain.
The greater the ignorance, the greater the dogmatism.

Attributed to William Osler, 1902

ChuckG
Posts: 99
Joined: Thu 17 Mar 2011 23:45
Location: Berkeley

Re: Muliple-Strain Bb Infections: Implications

Post by ChuckG » Tue 13 Aug 2013 23:46

Study Figure 2 for a visual demonstrating why IGeneX uses B31 and 297.

http://dc303.4shared.com/doc/XG86xab5/preview.html

dlf
Posts: 294
Joined: Sun 7 Apr 2013 15:36

Re: Muliple-Strain Bb Infections: Implications

Post by dlf » Wed 14 Aug 2013 1:43

I think LHCTom has given a wonderful summation for the questions Duncan was asking and Chuck G provided an excellent link for the Igenex testing and why two strains work better than one to be more sensitive for Western Blot testing.
Duncan also wrote,Can you get labs to break out the specific strains one has? I have had many tests for Lyme. The vast do not provide data that specific, i.e., which strain(s) you are infected with. That being said, I also have tests which DO seem to specify the strain. Some seem to indicate multiple strains, and even break out each strain's specific bands when they involve WB's. Maybe I am misreading them, but I don't believe so.
If it hasn't been made quite clear yet, those specific bands which appear on the tests Duncan is looking at relate to different proteins all contained in a SINGLE strain B31, or in the case of Igenex, the proteins contained in each of the two strains that they use together and report as a single test.(As shown in ChuckG's link)
Duncan wrote, Does it even matter? What are the ramifications of being infected with more than one strain? The first paper I found that even addresses the possibility of multiple strain infections was from 1999 and included Dattwyler. Sorry, no link. But, would being infected by multiple strains make it more difficult for one's body to clear the infections? Does treatment get a little trickier potentially? Or, will a conventional dose of abx take care of the whole bunch in a single collective swoop? Is there even a protocol for multiple strain infections? Or, to get darker and more speculative, if left untreated, could each strain develop unto its own peculiarities, and engender symptomalogies that are strain-specific, and become individually entrenched, and, well, you get the idea. It's the what-happens-if-you-let-one-strain-go-untreated scenario, potentially, times two. Or three.
As LHCTom pointed out, so far we really don't have enough understanding to know what the real world differences in having multiple infections mean in terms of symptoms, how they might be expressed over time and the best treatment for each of the various strains or combinations of strains. To give Duncan a little bit more reading on why strain, or more specifically the Outer Surface Protein C of a strain, can make a difference I would suggest

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC116539/

Four Clones of Borrelia burgdorferi Sensu Stricto Cause Invasive Infection in Humans
Gerald Seinost,1 Daniel E. Dykhuizen,2 Raymond J. Dattwyler,1,* William T. Golde,1 John J. Dunn,3 Ing-Nang Wang,2 Gary P. Wormser,4 Martin E. Schriefer,5 and Benjamin J. Luft1

And for some reading about multiple infections with different European species of Borrelia, posted by Panda on another thread:

http://www.lymeneteurope.org/forum/view ... 7&start=10

Mixed infections transmitted by Ixodes ticks in north-western region of Russia (clinical features, diagnosis, treatment)
Uskov, Alexander
Thesis, Moscow 2005

Another consideration which Duncan has not added into his mix of questions is that several other co-infecting pathogens like Babesia, Bartonella, Anaplasma phagocytophilum, etc.... which may be transmitted with the same tick bite are also very likely to change the nature and evolution of a persons illness and affect what treatments may be most useful.

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

Re: Muliple-Strain Bb Infections: Implications

Post by LHCTom » Wed 14 Aug 2013 4:26

I had positive Western Blots done at IGenex and it did not show the 2 strains separated out. I was not aware they offered this dual Western Blot clinically. It makes sense they would prepare both separately for their study but I have never heard of it being offered outside research and sent to a patient. Do they? In any case, these 2 strains are only two of dozens in the US.

http://www.igenex.com/labtest.htm


It appears to help as I was C6 positive at 3 different labs, Quest, Stony Brook and Igenex. They all use the same Immunetics kit which was tested by Wormser et al.

http://www.ncbi.nlm.nih.gov/pubmed/18724824

http://www.ncbi.nlm.nih.gov/pubmed/23062467
Effect of Borrelia burgdorferi genotype on the sensitivity of C6 and 2-tier testing in North American patients with culture-confirmed Lyme disease.
Wormser GP, Liveris D, Hanincová K, Brisson D, Ludin S, Stracuzzi VJ, Embers ME, Philipp MT, Levin A, Aguero-Rosenfeld M, Schwartz I.
Source

Division of Infectious Diseases, Department of Medicine, New York Medical College, Valhalla, NY 10595, USA. gary_wormser@nymc.edu
Abstract
BACKGROUND:

A potential concern with any serologic test to detect antibodies to Borrelia burgdorferi is whether the epitopes incorporated in the test provide sufficient cross-reactivity to detect infection with all of the pathogenic strains of the species. This is a particular concern for the C6 test, which is based on reactivity to a single peptide.
METHODS:

C6 testing and 2-tier testing were performed on acute-phase serum samples obtained from >158 patients with erythema migrans for whom the genotype of the borrelial isolate was defined on the basis of an analysis of the 16S-23S ribosomal DNA spacer region and/or on the genetic variation of the outer surface protein C gene (ospC). The sonicated whole cell-based enzyme-linked immunosorbent assay, the immunoblots used in the 2-tier testing, and the C6 assay all used antigens from B. burgdorferi sensu stricto strain B31.
RESULTS:

The sensitivity of C6 testing (69.5%) was greater than that of 2-tier testing (38.9%) (P<.001); the difference in sensitivity, however, was statistically significant only for patients infected with 2 of the 3 ribosomal spacer type-defined genotypes. The lower sensitivity of 2-tier testing was attributable to the low sensitivity of the immunoblot tests, rather than the first-tier enzyme-linked immunosorbent assay. There was also a trend for the sensitivity of 2-tier testing to vary according to the ospC genotype for the 14 genotypes represented in the study (P=.07); this relationship was not observed with C6 testing.
CONCLUSIONS:

Lack of sensitivity of the C6 test because of strain diversity seems less likely to be a limitation of this serologic test, compared with 2-tier testing in North American patients with early Lyme disease.
The greater the ignorance, the greater the dogmatism.

Attributed to William Osler, 1902

phyfe
Posts: 167
Joined: Sat 15 Sep 2012 19:28

Re: Muliple-Strain Bb Infections: Implications

Post by phyfe » Wed 14 Aug 2013 15:05

Since some people get the erythema migrans rash and some don't, could the strain be the defining criteria?

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

Re: Muliple-Strain Bb Infections: Implications

Post by LHCTom » Wed 14 Aug 2013 19:17

Since some people get the erythema migrans rash and some don't, could the strain be the defining criteria?
Yes, in part. There are many studies in process looking at pathogenic strategy and virulence factors in Borrelia. If you go to the NIH Categorical Spending - Lyme

http://report.nih.gov/categorical_spend ... %20Disease

Select one "Project Number" link

Then "Select Similar Projects" link

It will generate a list that includes all NIH funded Lyme studies including 2013. The regular categorical listing does not yet show 2013 studies. Then you can look through 2013 studies for ones looking at pathogenic or virulence and it will provide a look at summaries including hypothesis about what factors control virulence etc.. which in turn is related to symptoms observed in an infection. In general it how various genes are expressed that creates the factors for virulence etc.. Different strains can carry varied genes or plasmids that express differently. Those differences lead to how an infection will progress.

One study example for 2013 by Steven Norris is an attempt to "identify every B. burgdorferi gene that is important in the infection process"

http://projectreporter.nih.gov/project_ ... 622&icde=0

Lyme disease, the most common arthropod-borne disease in the United States, is caused by the spiral-shaped bacterium Borrelia burgdorferi and related organisms. These bacteria are transmitted by ticks and cause a long-term infection in people that affects the skin, nervous system, joints, and heart. Because we know so little about how B. burgdorferi causes disease, it is difficult to design better ways to prevent, diagnose, and treat Lyme disease. The goal of this study is to identify every B. burgdorferi gene that is important in the infection process, so that we can use the resulting information to help reduce the impact of Lyme disease on people in the United States and in other parts of the world.


And another point of view to study:

http://projectreporter.nih.gov/project_ ... 545&icde=0
The greater the ignorance, the greater the dogmatism.

Attributed to William Osler, 1902

ChuckG
Posts: 99
Joined: Thu 17 Mar 2011 23:45
Location: Berkeley

Re: Muliple-Strain Bb Infections: Implications

Post by ChuckG » Thu 15 Aug 2013 1:27

LHCTom wrote:I had positive Western Blots done at IGenex and it did not show the 2 strains separated out. I was not aware they offered this dual Western Blot clinically. It makes sense they would prepare both separately for their study but I have never heard of it being offered outside research and sent to a patient. Do they? In any case, these 2 strains are only two of dozens in the US.
Only a combined B31/297 WB is offered.
Comparison of specificity and sensitivity of IGeneX Lyme Western Blots using IGeneX criteria and CDC criteria for a positive Western Blot
Townsend Letter for Doctors and Patients, April, 2007 by Jyotsna S. Shah, Iris Du Cruz, Danuta Wronska, Steven Harris, Nick S. Harris

snip
B. burgdorferi culture and antigen preparation:

B. burgdorferi strains B31 and 297 were cultured at 34[degrees]C in Barbour-Stoenner-Kelly (BSK II) medium supplemented with 6% rabbit serum and 7.5% gelatin. (4) Cells were harvested at density [10.sup.7]-[10.sup.8] cells/ml, washed in PBS, and sonicated. Sonicated B31 and 297 cell lysates were pooled (1:1 by weight) together to ensure optimal band composition. Protein concentration was determined by optical density.

Pandora
Posts: 252
Joined: Tue 20 Mar 2012 14:58

Re: Muliple-Strain Bb Infections: Implications

Post by Pandora » Thu 15 Aug 2013 12:07

http://www.besjournal.com/Articles/Arch ... 85328.html
Optimization of Pulse-Field Gel Electrophoresis for Borrelia burgdorferi Subtyping*

GENG Zhen1,2,+, HOU Xue Xia1,+, HAO Qin1,+, ZHOU Hai Jian1, WANG Feng2,#, and WAN Kang Lin1,#
1.State Key Laboratory for Communicable Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China; 2. Department of Infectious Disease, The First Hospital, Jilin University, Changchun 130021, Jilin, China

Full-Text PDF: 584-591.pdf
---------------------------
As you know China has TWICE said you only need ONE band to say pos. because of ITS antigenic variation.
2010 Oct Abstract
http://www.ncbi.nlm.nih.gov/pubmed/21112481
2013
http://www.ncbi.nlm.nih.gov/pubmed/23425802
--------------------
http://www.canlyme.com/wb.html
The CDC criteria for a positive WB are as follows:

* For IgM, 2 of the following three bands: OspC (22-25), 39 and 41.

* For IgG, 5 of the following ten bands: 18, OspC (22-25), 28, 30, 39, 41, 45, 58, 66 and 83-93.
While removing bands 31-34.
Can we now estimate how many patients have been left to die with their syndrome of unknown origin because of the lies and failures?

dlf
Posts: 294
Joined: Sun 7 Apr 2013 15:36

Re: Muliple-Strain Bb Infections: Implications

Post by dlf » Thu 26 Feb 2015 22:42

I am hoping someone on this forum might be able to help out. Back in 2013, ChuckG posted the following link to:
Shah, JS et al. Improved Clinical Sensitivity for Detection of Antibodies to Borrelia burgdorferi Western Blots Prepared from a Mixture of Two Strains of B. burgdorferi 297 and B31, and Interpreted by In-house Criteria. This link used to work, but no longer does! If anyone can help with a new link (or a full text copy of this article), it would be very appreciated! If someone can help with a copy, please PM me, if anyone has a new link, it would be wonderful if you could post it.

Thanks!
Re: Muliple-Strain Bb Infections: Implications
Postby ChuckG » Tue 13 Aug 2013 22:46

Study Figure 2 for a visual demonstrating why IGeneX uses B31 and 297.

http://dc303.4shared.com/doc/XG86xab5/preview.html
ChuckG

Posts: 99
Joined: Thu 17 Mar 2011 23:45
Location: Berkeley

Post Reply