Archive for the ‘Testing’ Category

New Lyme Disease Test Distinguishes Between Early and Late-Stage Disease

ihttps://medicalxpress.com/news/2021-04-lyme-disease-distinguishes-early-late-stage

New Lyme disease test distinguishes between early and late-stage disease

April 7, 2021

by Frontiers

Lyme disease

For those who live in an area blighted by ticks, the threat of Lyme disease can cast a shadow over the joy of spring and summer. These blood-sucking arachnids can transmit bacteria into the bloodstream of their unsuspecting host, causing the disease. Early treatment is essential, but current tests are not usually sensitive enough to detect the disease in early-stage patients. A recent study in open-access journal Frontiers in Microbiology reveals a new test for Lyme disease, which is the first to reliably distinguish between early- and late-stage patients. The test detects a genetic sequence left by a virus that resides in Lyme-causing bacteria, and can detect just one bacterial cell in a small blood sample.

As the most common tick-borne infection, Lyme affects nearly 500,000 people in the U.S. every year. Symptoms include fever, fatigue, joint pain, and a distinctive ‘bullseye’ rash, but if left untreated, the disease can cause paralysis and even death. As such, is important, but difficult.

“Early diagnosis of Lyme disease is absolutely vital in reducing suffering, because early Lyme can be treated, but late Lyme is very difficult to treat,” explained Dr. Jinyu Shan of the University of Leicester, lead author on the study. “Current tests cannot typically detect the low numbers of bacteria in early-stage patient blood samples. Our goal was to design a highly sensitive to help doctors to identify Lyme disease as early as possible.” (See link for article)

______________________

Study here: https://madisonarealymesupportgroup.com/2021/03/31/targeting-multicopy-prophage-genes-for-the-increased-detection-of-borrelia-burgdorferi-sensu-lato-the-causative-agents-of-lyme-disease-in-blood/

Evidently the test is unique in that it is based on prophages that have a genetic sequence inserted into the bacteria by a virus which can escape the bacteria, and is more likely to be picked up in the blood due to having multiple copies within cells.

They found the test is sensitive and can detect one bacterial cell in .3 mL of blood.  Infected patients have between 1-100 bacterial cells per mL of blood.

The test is the first to distinguish between healthy, early-stage, and late-stage samples.

I must admit that the debacle with PCR testing for COVID has made me extremely skeptical and wary.  I hope that this isn’t too good to be true.  Time will tell.

Category:

Activism, Lyme, research, Testing

Current Serology – No Better Than A Coin Toss

https://www.change.org/p/the-us-senate-calling-for-a-congressional-investigation-of-the-cdc-idsa-and-aldf

Current serology is no better than a coin toss

MAR 31, 2021 — 

Please see the letter below addressed to the Chair of the NH Lyme Disease Study Commission in reference to a presentation from our last Zoom meeting.

You can hear/see my arguments at 1:01 in the recorded YouTube video link below.

———- Original Message ———-
From: CARL TUTTLE <runagain@comcast.net>
To: William Marsh <wmarshmd@gmail.com> CHAIR
Cc: All members of the study group
Date: 03/31/2021 8:59 AM
Subject: Topic for discussion at our next meeting (Dr. Martin’s Presentation)

I would like to add the following letter of concern as a topic for discussion at our next meeting and have copied all committee members so they can review the document prior to the meeting. 

Please reserve any discussion of these documents until our next scheduled Zoom meeting.

To members of the Lyme Study Commission,

Dear Rep Marsh,
I have some serious concerns regarding Dr. Martin’s informative presentation last Friday.

In reference to the 2013 C6 Elisa paper from Dr. Wormser, [1] this test is not widely used here in the US for the detection of Borrelia infections. When performing a search on the Quest Diagnostics website, I could not find the C6 Elisa listed as an option for Lyme disease.

Furthermore, test performance from that 2013 Wormser paper and 2018 pegalajar-jurado et al publication [2] was derived through the use of serum samples from patients who had the positive marker for Lyme; erythema migrans (bulls-eye rash) so testing results for the newer serology were positive because they were already positive. There was no performance data for those who never developed the bulls-eye rash. Once again this is misleading to the reader and highly inaccurate in real-world application. (what front line physicians are experiencing) Maine Department of Health is reporting incidence of rash at only 50%. So, half of the patient population is not being represented in these studies. That is a serious concern.
 
To my knowledge, the Modified Two-Tiered Testing Algorithm (MTTTA) is not in use outside of Dr. Martin’s lab and irrelevant to our study commission at this time so any performance numbers offered by Dr. Martin are meaningless. Until such time that these newer tests are fully implemented (if ever) performance characteristics should not be acknowledged or posted in the minutes of the Lyme Study Commission.

Although current testing methods are evolving, we are still using the conventional WCS ELISA–immunoblot-based 2-tier testing algorithm. Dr. Durand (ILADS physician) and I have provided documentation supporting the fact that current serology is no better than a coin toss and this fact must be shared with the public along with the fact that humans do not produce antibodies for 4-6 weeks after a tick bite.

Here are the missing disclaimers:

1. “Health-care providers are reminded that a diagnosis of Lyme disease should be made after evaluation of a patient’s clinical presentation and risk for exposure to infected ticks, and, if indicated, after the use of validated laboratory tests.

2. “This surveillance case definition (two of three IgM and Five of ten IgG bands) was developed for national reporting of Lyme disease; it is not intended to be used in clinical diagnosis”

Respectfully submitted,

Carl Tuttle
Hudson, NH

References

1. Single-tier testing with the C6 peptide ELISA kit compared with two-tier testing for Lyme disease
 https://demystifyingmedicine.od.nih.gov/dm13/2013-03-19/dmid-v75-y2013-p9.pdf

2. Evaluation of Modified Two-Tiered Testing Algorithms for Lyme Disease Laboratory Diagnosis Using Well-Characterized Serum Samples
 https://jcm.asm.org/content/56/8/e01943-17

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For more:

There has been a concerted effort by ‘authoritie’s against direct testing. 

The simple reason for this is our public health ‘authorities’ have patents on Lyme testing.

They also want us to take their controversial Lyme vaccines which would only work if Lyme disease does not cause chronic illness – hence the denial of persistent infection.  Our public authorities also have conflicts of interests regarding these vaccines.

The CDC is not to be trusted at all.  

We need to stop working with these agencies and raising money for their flawed, biased science that continues to abuse and neglect sick patients.

 

Category:

Activism, Lyme, Testing

Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato, the Causative Agents of Lyme Disease, in Blood

https://www.frontiersin.org/articles/10.3389/fmicb.2021.651217/full

ORIGINAL RESEARCH ARTICLE

Front. Microbiol., 15 March 2021 | https://doi.org/10.3389/fmicb.2021.651217

Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato (s.l.), the Causative Agents of Lyme Disease, in Blood

Jinyu Shan1*†, Ying Jia1†, Louis Teulières2, Faizal Patel1and Martha R. J. Clokie1*
  • 1Department of Genetics and Genome Biology, University of Leicester, Leicester, United Kingdom
  • 2PhelixRD Charity 230 Rue du Faubourg St Honoré, Paris, France

The successful treatment of Lyme disease (LD) is contingent on accurate diagnosis. However, current laboratory detection assays lack sensitivity in the early stages of the disease. Because delayed diagnosis of LD incurs high healthcare costs and great suffering, new highly sensitive tests are in need. To overcome these challenges, we developed an internally controlled quantitative PCR (Ter-qPCR) that targets the multicopy terminase large subunit (terL) gene encoded by prophages that are only found in LD-causing bacteria. The terL protein helps phages pack their DNA. Strikingly, the detection limit of the Ter-qPCR was analytically estimated to be 22 copies and one bacterial cell in bacteria spiked blood. Furthermore, significant quantitative differences was observed in terms of the amount of terL detected in healthy individuals and patients with either early or late disease. Together, the data suggests that the prophage-targeting PCR has significant power to improve success detection for LD. After rigorous clinical validation, this new test could deliver a step-change in the detection of LD. Prophage encoded markers are prevalent in many other pathogenic bacteria rendering this approach highly applicable to bacterial identification in general.

Introduction

Lyme disease (LD) is the most common tick-born disease with approximately 476,000 patients in the United States annually during 2010–2018 (Kugeler et al., 2021). LD is caused by a group of bacteria classified together as the Borrelia burgdorferi sensu lato (s.l.) complex, that comprises a clade of more than 20 species including B. burgdorferisensu stricto (s.s.) which dominates in United States, and B. garinii and B. afzelii which are prevalent in Europe and Asia. The LD-causing bacteria are generally transmitted to humans after they are bitten by ticks of the Ixodes family infected with LD causing Borrelia. However, recent reports have raised concerns over Borrelia transmission through blood transfusion based on observations that Borrelia can survive and circulate in the human bloodstream (Pavia and Plummer, 2018).

Currently, LD diagnosis is based on the overt clinical manifestation of disease in the form of erythema migrans (EM) skin lesions, commonly known as a ‘bull’s-eye’ rash and a history of tick exposure. Although EM lesions occur in 70 to 80% of infected individuals, only a third of these patients develop the classic ‘bull’s-eye’ rash, and many other types of skin lesions can occur which are easily confused with EM (Chaaya et al., 2016). In addition to the EM uncertainty, other common symptoms of LD such as fatigue, muscle pain, headache, and perceived cognitive dysfunction largely overlap with an array of other diseases, including other tick-borne diseases. One such example is Relapsing Fever (RF), which is caused by close relatives of the LD-causing bacteria, such as Borrelia miyamotoi(Wormser et al., 2019). The two Borrelia ‘groups’ responsible for LD and RF have caused great concern and clinical confusion, as they are morphologically similar and present with almost indistinguishable clinical symptoms (Bergström and Normark, 2018). Despite this, they respond to different antibiotics and treatment regimens (Koetsveld et al., 2017). Another example of confusion surrounding LD is the co-infection caused by Bartonella spp. This genus of bacteria is emerging as an increasingly common human infection (Anderson and Neuman, 1997). Much of the controversy surrounding LD and co-infections with Bartonella and/or B. miyamotoi is due to the lack of a reliable and sensitive diagnostic method to detect and distinguish between the three groups of bacteria, the LD and RF causing Borrelia and Bartonella (Schutzer et al., 2019). Therefore, laboratory tests to determine and distinguish between LD and co-infections play a vital role in the correct diagnosis and consequent treatment with different antibiotics.

Scientists have faced several challenges with LD detection including patients presenting with a delayed antibody response and a low number of Borrelia cells typically found in human clinical samples (Moore et al., 2016). Although it is particularly difficult to diagnose LD early, it is critical, as it is far easier to treat the disease when it is detected at an early stage (Theel et al., 2019). Bacteria-targeting approaches, such as polymerase chain reaction (PCR) detecting the Borrelia chromosomal DNA, can potentially identify early LD but is relatively insensitive detecting only between 30-50% of positive cases, and is therefore deemed to have little clinical utility (Schutzer et al., 2019). The reasons behind the poor sensitivity of the current PCR methods in Lyme detection are twofold; first, the current PCRs target Borrelia genomic DNA regions that have only one copy in each bacterium, such as the bacterial 16S rRNA gene, RecA gene, and the 5S-23S intergenic regions (Brettschneider et al., 1998; Liveris et al., 2012; Waddell et al., 2016; Lohr et al., 2018; Schutzer et al., 2019). Second, at least some Borrelia species are ‘tissue-bound’ and are only transiently found circulating in the blood (Liang et al., 2020).

In response to these diagnostic challenges, we adopted a novel approach, taking advantage of the fact that most pathogenic bacteria carry multiple complete or partial prophages (phages associated with bacteria) (Argov et al., 2019). These prophage sequences can form the bases of a template from which quantitative PCR (qPCR) primers and probes can be designed. It is known that Borrelia carry a large number of linear and circular plasmids (comprising between 33-40% of the Borrelia genome), among which the cp26 and cp32, and the lp54 linear plasmid, are evolutionarily stable (Casjens et al., 2017). Of these paralogous plasmids, cp32 has been experimentally determined to be a Borrelia burgdorferi prophage thus it is highly likely that many of its homologs are also prophages (Eggers and Samuels, 2000).

In this paper we have demonstrated for the first time in Borrelia-related diagnostics that it is possible to overcome the sensitivity challenges associated with LD detection. We highlight the potential of our test to discriminate between healthy volunteers, early LD, and late LD patients. We present data from a systematic and comprehensive study that evaluate the use of the multicopy phage terminase large subunit (terL) gene as a molecular marker for the detection of Borrelia species. The analytical performance of the terL-targeting qPCR (referred to as Ter-qPCR) was thoroughly evaluated, and the test was shown to be able to detect one single Borrelia cell from blood samples. The diagnostic potential was evaluated using a set of blood and serum samples collected from healthy volunteers and individuals who were clinically diagnosed with LD.

In summary, we demonstrate that a quantitative phage-based PCR has the potential to change the diagnosis of LD from blood samples. This approach of detecting bacteria-specific phages may be applicable to infections other than LD such as sepsis caused by Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa etc. (Minasyan, 2019), as long as suitable phages are identifiable.

Results and Discussion

Each Borrelia species has a distinct amount of species specific variation in its prophage sequences; thus these prophages can be used as a proxy to identify the bacteria because of the tight correlation between them and the exact prophages found in each Borrelia host. As there are multiple prophages per Borrelia cell, the detectable signal is higher for prophages than bacteria. Furthermore, evidence suggests that Borrelia prophages can be released outside the Borrelia cells following encounters with stressors such as antibiotics (Eggers and Samuels, 2000). In this study, we confirmed that Borrelia prophages can escape from the bacterial host cell in a spontaneous manner. Taking advantage of the multicopy and free movement of Borrelia prophages, the approach to target prophages instead of bacteria will bypass the cryptic and tissue-bound feature that typifies human Borrelia infections (Liang et al., 2020). Thus, we have a greater chance of detecting the prophages in blood even when the bacteria may not be present or present in extremely low numbers. In this sense, prophages are somewhat analogous to Borrelia ‘footprints’.

(See link for full article)

_______________________

**Comment**

Important excerpt:

Borrelia Strains and Cultures

The Borrelia strains used in this study are listed in Table 1.

  • Ten strains were provided by Professor Sven Bergström, Department of Molecular Biology, Umea University, Sweden. Seven strains were purchased from the Pasteur Institute and DSMZ (German Collection of Microorganisms and Cell Cultures GmbH).
  • Two strains were provided by the Center for Disease Control and Prevention (CDC), United States, and
  • two by Cecilia Hizo-Teufel from the German National Reference Centre for Borrelia. 

For more: 

Category:

Lyme, research, Testing

Study of a Potential Test for Persistent Lyme Disease

https://flightpath.bio

FLIGHTPATH BIOSCIENCES’

Study of a Potential Test for Persistent Lyme Disease

Thanks to the enthusiastic response from the Lyme community, this study has met its current enrollment goal. Therefore, Flightpath Biosciences’ Study of a Potential Test for Persistent Lyme Disease will not be accepting any more applications at this time.

We’ll keep you posted as the project proceeds or if we reopen the study for further enrollment.

For questions related to this study, please contact: FlightpathLymeStudy@gmail.com

https://news.northeastern.edu/2020/09/29/intestinal-bacteria-could-give-doctors-an-objective-test-for-chronic-lyme-disease

Excerpt:

Fatigue, muscles aches, brain fog—are these symptoms of chronic Lyme disease, or merely side effects of the daily grind of human existence? It’s hard to tell. 

Chronic Lyme disease, also known as post-treatment Lyme disease syndrome or PTLDS, is incredibly hard to diagnose because symptoms vary greatly, and there is currently no biological test to detect the disease.

Now, Kim Lewis, University Distinguished Professor of biology and director of the Antimicrobial Discovery Center at Northeastern, has proposed a new way to objectively diagnose this elusive disease by analyzing the microbes in a patient’s gut. 

(Go to link for article)

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**Comment**

Lewis states there are about 800,000 people in America living with PTLDS.  I have written before about this confusing moniker that it means different things to different people.  For instance, microbiologist Holly Ahern states there are two groups of patients: those diagnosed and treated early and those diagnosed and treated late.  The PTLDS label only concerns the first group and only represents about 10-20% of people going on with persistent symptoms.  These low percentages are typically what researchers are referring to.  The label leaves out a much larger group (30-40%) that is diagnosed and treated late.  

This second group represents nearly all the patients I work with that never gets addressed by research because their cases are sticky, hard to define, and by nature don’t fit well into a research study design.

According to Lewis, people with PTLDS have an abundance of a type of bacteria called Blautia and a suppression of a type of bacteria called Bacteroides (which explains why Lyme/MSIDS patients suffer with inflammation, digestion, improper immune responses, depression and anxiety).

Unfortunately, this bacterial disregulation is also seen in many other diseases.

Flightpath is also working on making an oral form of the antibiotic azlollicin available, which has demonstrated in vivo efficacy in mice by significantly inhibiting the growth of drug-tolerant Borrelia burgdorferi (Bb) bacteria better than doxycycline (the standard of care), and reducing inflammation.

 

Category:

Gut Health, Inflammation, Lyme, research, Testing

COVID Long Haulers & Lyme Disease

https://globallymealliance.org/long-haul-covid-19-brings-attention-to-persistent-lyme/

by Jennifer Crystal

This past January, the New York Times published an article entitled “What if You Never Get Better From Covid-19?”. Many Lyme patients sent it to me in frustration. “This is just like Lyme disease!” they exclaimed. “We haven’t gotten better, either. Why is everyone talking about COVID long-haulers when we’ve been suffering for years?”

I hear that frustration. I also think it’s precisely this attention on long-haul COVID that will finally shed light on, and bring credibility to, persistent Lyme disease.

In fact, the article itself draws comparisons between COVID-19 and Lyme disease, as well as myalgic encephalomyelitis, or chronic fatigue syndrome (ME/CFS). As with Lyme disease, the cause of ongoing COVID-19 symptoms is now under investigation: is it ongoing infection? Is it a post-acute syndrome? Is it an immune response? These are questions that have echoed in the Lyme world for years, causing polarizing debate that leaves some patients fighting for validation, diagnosis, and treatment.

Now, that struggle is coming out of the echo chamber and into the limelight—pun intended—in a world that has newfound understanding of long-term illness and its effects. Words like “immunocompromised” and “long-hauler” were not part of the general lexicon before COVID-19, even though they’ve been part of Lyme patients’ vocabulary for years. Now these words are vernacular, and that’s good news for Lyme patients, ME/CFS patients, and anyone else struggling with a complex illness that strikes different patients in different ways. As the article states, “The Lyme problem is underrecognized but immense.” One silver lining of a horrific pandemic that has killed half a million Americans and left countless others still struggling is that it’s bringing recognition to illnesses that have long caused long haul suffering.

Indeed, just a few weeks after the aforementioned article was published, New York Times Opinion Columnist Ross Douthat wrote “Long-Haul Covid and the Chronic Illness Debate: What persistent Covid cases might have in common with chronic fatigue syndrome and Lyme disease, and why it matters”. A Lyme patient himself, Douthat wrote, “Living through the coronavirus era after spending so many years in the world of Lyme disease is a strange experience because you can see all kinds of different pieces of the tick-borne epidemic refracted strangely in the Covid pandemic—disputes over testing, mysterious and shifting symptomatology, expert failures and medical populism, and controversies around what it means when the disease just hangs around indefinitely.”

Even back in spring 2020, when COVID-19 testing difficulties were at their height, people were taking notice of the similarities to Lyme disease. I was interviewed on this topic for an article on Undark.org called “In the Uncertainties of Lyme Testing, Lessons for COVID-19”.

One important distinction that Douthat notes between long-haul COVID and Lyme, ME/CFS and other illnesses is that “we’re taking the lived experience of long-haul [COVID] patients seriously—probably because we have so many of them all at once—instead of treating them as weaklings or hypochondriacs.”

To that, I can speak personally. For years before I was diagnosed with Lyme disease, babesiosis, and ehrlichiosis, I fought just for people to believe that I was sick; that my flu-like fatigue, insomnia, hallucinogenic nightmares, trembling hands, difficulty concentrating, systemic hives, and fevers were not psychosomatic. Then, once I was properly diagnosed, I fought for understanding from people who literally did not believe in my illness—as if a documented physical disease is simply an opinion or mystical power one can choose to believe in or not.

Conversely, when I first developed COVID-19 symptoms in March 2020, not one person questioned whether I actually had COVID-19, even though I had a (false) negative test. No one said, “Oh, you must have something else” or, “It must just all be in your head.” They knew that my shortness of breath, dry cough, low-grade fever, and loss of taste and smell meant COVID-19. When the positive antibody test came back in June, it was simply validation of what everyone already knew to be true.

When I had COVID-19, I only had to fight to get well, not to be believed.

What a difference that validation meant, as I discussed in my “Comfort in Solidarity” post. I want that type of understanding for all Lyme patients, in all stages of illness. I believe the COVID-19 pandemic will help get us there.

GLA is offering a free webinar, “COVID-19 Vaccines and Lyme Patients: What You Need to Know” on March 3rd at 6:00 p.m. You can register here.

For more blogs, click here

jennifer crystal_2

Opinions expressed by contributors are their own.

Jennifer Crystal is a writer and educator in Boston. Her memoir about her medical journey is forthcoming. Contact her at lymewarriorjennifercrystal@gmail.com.

 

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**Comment**

She was ill with something, but it is uncertain if she truly had COVID based on two things: it’s never been isolated and proven to cause disease, as well as the fact antibody as well as PCR testing is as bad if not worse than Lyme.  How can you have something that hasn’t been proven to exist and cause illness?  

This is truly the foundational issue that most are bypassing.  Our public ‘authorities’ have been getting away with this forever.

There is an inaccuracy in the Times article which states there are 329,000 people infected with Lyme per year. The CDC just upped that number to 476,000.

There were; however, important issues exposed by the Times article, but I don’t hold the same optimism Crystal does. If our public ‘authorities’ can bamboozle nearly the entire American populace with a ‘virus mania’ dating back decades, and convince them to take the jab on an experimental, fast-tracked injection that hasn’t been proven to be safe or effective, doesn’t stop transmission or infection, changes your DNA, can monitor and track you, and is said by whistleblowers to cause sterility, has caused 40 times more deaths in Israeli elderly and 240 times more deaths in the younger ages in the 5 week vaccination period than the virus itself, and the death of 25% of German nursing home residents, I highly doubt this same populace will embrace the complexity of Lyme/MSIDS.

I’d love to be proven wrong.

In my experience, the only forward progress we’ve made has been when we’ve done the work ourselves with independent researchers unaffiliated with government funding.

Category:

Activism, Lyme, Testing, Viruses