@Sproetje, heb je het Opinie artikel goed gelezen en de inhoud goed begrepen? En welke collega in België bedoel je? De 'Phelix Phage Borrelia test' werd zelfs gepromoot in Nederland bij de ZonMw - Werksessie ME/CVS 4 november 2020 Utrecht; Bron https://www.zonmw.nl/nl/actueel/nieuws/ ... ies-mecvs/ De clinicus is een collega van Mijatovic van Red Laboratories. Clinicus stelt zich drie vragen; Bron https://www.zonmw.nl/fileadmin/zonmw/do ... irleir.pdfSproetje schreef:Misschien kan J. Hovius al de vragen die hij heeft eens stellen bij zijn collegea in België
Is per slot van rekening vlak naast de deur.
En wetenschappers onderling info uitwisselen zou naar mijn bescheiden mening iedereen ten goede komen
Het is niet het lab van Wuhan, waar niemand binnen kan komen
In 2020 wordt gesteld dat me/cvs patiënten B. miyamotoi/Relapsing Fever hebben en de 'Phelix Phage Borrelia test' een 'nieuwe doorbraak' zou zijn - 'Bacteriophage Lyme Test Offers ME/CFS Patient New Possibility'; Bron https://www.healthrising.org/blog/2020/ ... eriophage/ De diagnose wordt gesteld met de 'Phelix Phage Borrelia test'. De patiënte is tot nu toe niet verbeterd met de behandeling en is nog steeds bedlegerig; Bron https://twitter.com/domstanc..Latente actieve infecties?
1. Tot nu toe ongekende infecties.
50% Borrelia infectie (B. Miyamotoi, B. Recurrent fever type, B. Burgdorferi) – nieuwe onderzoekstechnieken (phage Borrelia qPCR)
Andere: Yersinia Pseudotuberculosis, Tularemie, andere gramnegatieve bacteriën, parasieten, enkele virussen zoals Parvovirus B19..
Het begint meer en meer op het 'xmrv/mlv-virus debacle' te lijken; Bron viewtopic.php?f=38&t=2435&p=27882#p27534
Veel me/cvs patiënten kregen een 'positieve uitslag' op 'xmrv/mlv virus' zonder het verstrekken van data gegevens door Red Laboratories.
Er ontstond veel ongerustheid over de 'positieve uitslagen xmrv/mlv virus' de me/cvs patiënten werd verteld dat zij kanker konden hebben of zouden gaan ontwikkelen. Er is door de patiënten veel geld betaald voor de testen.
Patiënten werden behandeld met een experimentele en kostbare behandeling bestaande uit verschillende soorten antibiotica, Nexavir (4ME/Kutipressin) en supplementen dat werd verkocht via een eigen onderneming en waar veel geld mee is verdiend. De me/cvs patiënten hebben met de behandelingen geen verbetering of herstel gekregen. Er zijn nooit excuses gemaakt en de mensen hebben geen geld terug gekregen.
De test bleek bij validatie en verificatie door negen andere geaccrediteerde medische laboratoria ondeugdelijk te zijn.
Het is medisch en ethisch onverantwoord dat de artsen en de therapeuten die met de 'Phelix Phage Borrelia test' werken bij hoopvolle en of wanhopige patiënten (met Lyme, chronische Lyme, ziekte me/cvs, mensen (kinderen) met Autisme (Autism Spectrum Disorders (ASD) en mensen zonder een diagnose) énkel op basis van de 'positieve uitslagen' van de 'Phelix Phage Borrelia test' een diagnose
B. miyamotoi/Relapsing Fever en of Borrelia burgdorferi sensu lato stellen en de patiënten een kostbare en experimentele behandeling bestaande uit een zware antibiotica behandeling c.f. in combinatie met disulfiram en of dapsone voorschrijven waar géén medische en wetenschappelijke onderbouwing voor is.
De zware antibiotica soorten c.f. die langdurig worden ingezet lijken op chemo. Is het verstandig dat zieke mensen dat in hun lichaam krijgen? Er worden tot nu toe geen getuigenissen van patiënten gedeeld in de media over verbetering of genezing.
Een verkeerd gestelde diagnose en een verkeerde voorgeschreven behandeling kan grote gevolgen hebben voor de gezondheid van mensen. Een verkeerde behandeling kan érg schadelijk zijn voor de gezondheid. Patiënten kunnen als gevolg van de behandeling ernstige langdurige (of blijvende) klachten (neurologische en of psychische) krijgen en of blijvende schade aan de darmen en de werking van het afweersysteem (immuunsyteem) krijgen. Mensen worden in dat geval niet beter maar zieker.
Recente beschrijvingen zijn:
Frontiers in Medicine
20 April 2020
'Potential Patient-Reported Toxicities With Disulfiram Treatment in Late Disseminated Lyme Disease' by Alain Trautmann, Hugues Gascan and Raouf Ghozzi; Bron https://www.frontiersin.org/articles/10 ... 00133/full
Lymedisease.org
23 november 2020
By Dr Daniel A. Kinderlehrer MD (internist) - Patients can respond very differently to disulfiram. Be cautious (over 100 patients); Bron https://www.lymedisease.org/kinderlehre ... m-caution/
Lymedisease.org
8 februari 2021
Kinderlehrer videos discuss disulfiram treatment, mast cell, and more; Bron https://www.lymedisease.org/kinderlehre ... mast-cell/
Lyme Disease.org
5 oktober 2021
LYME SCI: My re-cap of recent LDA/Columbia Lyme conference; Bron https://www.lymedisease.org/lda-columbi ... onference/
Initial Study - 'Disulfiram: A Test of Symptom Reduction Among Patients With Previously Treated Lyme Disease' by Brian A Fallon; Bron https://clinicaltrials.gov/ct2/show/NCT03891667On October 2, I attended the 21st annual scientific conference put on by the Lyme Disease Association and Columbia University’s Vagelos College of Physicians & Surgeons. The virtual event was entitled Lyme & Other Tick-Borne Diseases: Research for a Cure.
Disulfiram
..Dr. Fallon made a point of stating the use of disulfiram is experimental and not currently approved for Lyme disease..
De resultaten van de pilot-studie (voorlopige studie ter verkenning) met 24 personen zijn uitgesteld naar 31 maart 2022.
Amsterdam UMC; Bron https://www.amc.nl/web/mijn-afspraak/mi ... ntest-.htm en de Stichting Tekenbeetziekten; Bron https://www.tekenbeetziekten.nl/de-teek ... miyamotoi/ en Bron https://www.tekenbeetziekten.nl/de-teek ... /diagnose/ en de Biomaatschappij; Bron https://www.biomaatschappij.nl/artikel/ ... -maatwerk/ geven duidelijke informatie over B. miyamotoi en
de fagentest.
..Of de fagentest een Borrelia specifieke profaag detecteert is tot nu toe onduidelijk, want er wordt bij de test een PCR test gebruikt om een meer algemeen stukje erfelijk materiaal van bacteriofagen aan te tonen. Om die redenen is zowel ‘specificiteit' (dat het om de Borrelia bacterie gaat en welke) als ‘het aantonen van actieve infectie’ van deze test zeer dubieus. Zie ook de wetenschappelijke publicatie van 15 maart 2021.
De fagentest is een kostbare, nog weinig gangbare test en verkeert nog in een experimentele fase. Voor de Borrelia bacterie wordt deze test alleen aangeboden door een laboratorium in België en niet vergoed door zorgverzekeraars. De test is niet gevalideerd en er is een reëele kans op fout-positieve resultaten....Recentelijk krijgen wij veel vragen en verwijzingen over de zogenaamde ‘fagentest’ naar B. miyamotoi en B. burgdorferi..
..De fagentest staat dus nog in de kinderschoenen, maar wordt al wel via labs in het buitenland aan patiënten aangeboden. Het ontbreken van bewijs voor de betrouwbaarheid van de testuitslag wordt vaak niet of nauwelijks aan de patiënten medegedeeld..
..De fagentest kan dus wat ons betreft op dit moment dus niet helpen bij het stellen van de diagnose Lymeziekte of B. miyamotoi-ziekte..
Het is gezien alle bovenstaande informatie goed dat het Nederlandse team van wetenschappers&onderzoekers een Opinie artikel heeft gepubliceerd. De inhoud van het gepubliceerde Opinie artikel: 'Methodological Shortcomings in the Study on a Prophage-based PCR Test for Lyme Borreliosis' is meer dan duidelijk.
Frontiers in Microbiology..The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms..
Publicatie 15 maart 2021
ORIGINAL RESEARCH article - 'Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato (s.l.), the Causative Agents of Lyme Disease, in Blood' by by Jinyu Shan, Ying Jia, Faizal Patel, Louis Teulières and Martha R. Clokie; Bron https://www.frontiersin.org/articles/10 ... 51217/full
Het laboratorum Immunetics zit in Boston in de V.S...All participants were diagnosed by Dr. LT according to the ILADS (International Lyme and Associated Diseases Society) guidelines (Cameron et al., 2014). Some of individuals who did not recall a tick bite and had no LD-like symptoms (ILADS guidelines) served as healthy volunteers. All healthy volunteers were also negative to Lyme C6 ELISA that was performed according to manufacturer’s instructions (Immunetics: B. burgdorferi (Lyme) ELISA kit: DK-E352-096). A total of 312 samples (156 whole blood and 156 serum samples) were collected from 78 individuals between April-June 2017 (23 healthy volunteers with no ‘identifiable’ LD symptoms, 13 early stage and 42 late stage LD patients)..
Roxy schreef:Belangrijk nieuws over de 'Phelix Phage Borrelia test'. Belangrijk nieuws voor de mensen die met de 'Phelix Phage Borrelia test' zijn getest en een diagnose Borrelia burgdorferi sensu lato (s.l) en of B. miyamotoi/Relapsing Fever hebben gekregen.
Frontiers in Microbiology
OPINION article
Publicatie 13 december 2021
Opinion: 'Methodological Shortcomings in the Study on a Prophage-based PCR Test for Lyme Borreliosis' by Freek R. van de Schoor,
M. E. Baarsma, Mariska M. G. Leeflang, Volker Fingerle, Gabriele Margos, Joppe W. Hovius and Alje P. van Dam; Bron https://www.frontiersin.org/articles/10 ... 02131/fullWe read the article by Shan et al. (2021) with great interest, as new diagnostic tests for Lyme borreliosis (LB) are urgently needed (Cruickshank et al., 2018; Dessau et al., 2018). The article represents a proof of principle paper and an initial validation of an already commercially available test [Phelix Phage Borrelia—R.E.D. Laboratories (redlabs.be)]. We have various concerns regarding the study design, novelty of the approach, technical aspects of the assay, statistical analyses, and the conclusions, which must be addressed. Of note, several statements in the introduction are speculative and not supported by the references, but unfortunately, the word limit of our opinion does not allow us to elaborate on this.Gene sequences and PCR results
The concept of targeting genetic material from bacteriophages rather than from bacteria for clinical diagnosis is intriguing and—while not entirely new—it is still relevant today (Amouriaux et al., 1993). In a previous publication, Amouriaux et al. (1993) describe a similar approach targeting a plasmid region with sequence overlap to sequences used in the current publication (Figure 1A). However, the authors do not prove that bacteriophages are present and circulating in human blood. Therefore, the difference in sensitivity between the 16S PCR and terL PCR could actually be due to the difference in sensitivity between using a single-copy (16S) and a multi-copy (terL) target. This principle is well-known in bacteriology (Roosendaal et al., 1993). In addition, the genetic variation between and within the Borreliella burgdorferi sensu lato (s.l.) species of cp32 bacteriophage sequences is not discernible from the manuscript. In the alignment shown in Figure 2 in the article by Shan et al., the authors use cp32 genes of B. burgdorferi sensu stricto (s.s.)., but not of other species. B. burgdorferi s.s. B31 has the highest number of cp32 (n = 13) in comparison to 16S rRNA, a single copy chromosomal locus. We would be very interested to see how Borrelia afzelii and Borrelia garinii would have performed in analyses using spiked blood samples, as these are the most common genospecies causing clinical symptoms in Europe, but have—according to the authors' Table 1—fewer cp32 plasmids (n = 8, n = 4, respectively). In their paper, the geographical origin of patients is not described, but the authors state that “patients were diagnosed by Dr. LT,” referring to Louis Teulières, who is based in Paris, France, where—like in the rest of the European continent—LB is caused mainly by B. afzelii and B. garinii (Stanek et al., 2012). However, the test method is based on the terL gene derived from the North American strain of B. burgdorferi s.s. B31. Furthermore, the authors include extremely low-positive signals in their results. Of the 23 healthy controls, 21 showed a positive signal in the terL PCR in at least one of the 12 samples. Whereas incidental carriage of Borrelia-DNA in blood of healthy persons, as suggested by the authors, might occur, it is highly unlikely that this would be found in over 90% of the population. This strongly suggests that at least some low-positive results represent unspecific signals or signals which are a result of DNA cross-contamination.Selection Criteria LB Patients
Another concern is the patient selection and interpretation of the clinical data. The manuscript lacks any description of patient characteristics, and does not report inclusion or exclusion criteria. Absence of clear eligibility criteria may indicate selection bias. Criteria for patient selection in an LB-related diagnostic test accuracy study should be clear and unambiguous, for example based on European guidelines (Mygland et al., 2010; Stanek et al., 2011; Hofmann et al., 2017). The authors refer to the ILADS guideline (Cameron et al., 2014), which in itself does not contain any diagnostic criteria. Without unambiguous criteria, one cannot ensure that these individuals were in fact patients with LB (Stanek et al., 2011; Lantos et al., 2021). It is also unclear what is meant by “early LD” and “late LD.” Would Lyme neuroborreliosis (LNB) be classified as early or late LD, for example (Koedel and Pfister, 2017)? Were there any LNB patients at all? If so, how were they diagnosed?Statistical Analysis
We attempted to replicate the analyses presented by the authors in their Figure 7, using SPSS version 25. The authors describe having used Mann-Whitney U-tests to compare early LB patients, late LB patients, and healthy volunteers (HVs). However, they do not describe precisely how the results from the different groups have been compared. Their original dataset contains six test results on whole blood (WB) and six on serum for each participant, but it is unclear whether they analyzed all results, if they analyzed the mean per participant or used any transformation of the data. We replicated the Mann-Whitney U-tests to test for a difference between the different participant groups, using the mean values of the six iterations of each test per participant. While the authors' reported means and the means calculated by us were identical, our p-values were inconsistent with those reported by the authors. Subsequent analyses using other aggregate functions (such as medians) as input for our statistical tests did not result in p-values consistent with those reported by the authors either (data not shown). In contrast, when we used the six iterations of the terL assay per participant separately, as if they were independent values, the levels of statistical significance match those reported by the authors. By doing so the authors seem to have artificially inflated their statistical power by increasing their sample size six-fold. This may have resulted in identical mean values, but incorrect and much lower p-values. More so, the results from serum and WB samples from one participant are not independent, as both measurements were done in the same person. Therefore, a Wilcoxon Signed-Rank test would have been more appropriate to compare serum and WB within one patient group. The authors do not describe what statistical test they used, but if this was a Mann-Whitney U-test as described in the Methods section, then this is inappropriate.The Conclusions are Not Supported by the Data
The aforementioned considerations cast substantial doubt on the reliability of the results, but—when interpreted with caution—do not undermine the value of the authors' hypothesis. Unfortunately, the conclusions drawn by the authors from the results are inappropriate. The authors state that their assay can distinguish early LB, late LB, and HVs. These conclusions are not supported by the data.
The mean/median copy numbers may be significantly different at a group level—even though we have shown in this manuscript's Figures 1B,C and Supplementary Table 1 that they are not for most comparisons—but that does not imply diagnostic power. Only if there is little or no overlap between numeric values, will the assay be able to distinguish a patient from a non-patient. A simple scatterplot of the data shows there is a high degree of overlap between the groups. Subsequent ROC-analysis Figures 1C,D on the mean/median copy numbers shows that—when a minimally acceptable specificity of 90% is applied—the maximally attainable sensitivity is 62% (WB-MEAN: HV vs. early, cutoff at 1.275) or 57% (WB-MEAN: HV vs. late, cutoff at 1.283). This is worse than single-tier or modified two-tiered testing (MTTT) serology in EM and far worse than any type of serology in late LB (Leeflang et al., 2016; Waddell et al., 2016; Branda et al., 2017). Additional ROC-analyses show that the ability to discriminate between early vs. late LB is even (much) worse (data not shown). Please note that these analyses were performed with a small number of samples (early: n = 13; late: n = 42; HV: n = 23). It is much more likely that the assay lacks specificity and that many HVs had false-positive results, rather than suffer from asymptomatic B. burgdorferi infection, as the authors claim.
We must also point out that the manuscript suffers from flawed circular reasoning and over-interpretation. The fact that the groups differ with respect to the primary study parameter does not prove that they are LB patients or HVs. Participants' status as belonging to either group is the starting point for investigating potential differences in terL levels, not a conclusion that can be drawn when these groups are indeed shown to be different on this outcome. The authors postulate that their test could be used to monitor LB treatment outcomes, yet, this study does not report on any follow-up samples or treatment outcome to support this claim. They further state that the Ter-qPCR could be used to indicate which treatment option may work best, however, the choice of treatment option is not supported by any of the data in this article.Conclusions
We conclude that while this technique might be promising, the paper provides more questions than answers and contains a large number of inaccuracies. We would be interested to see the Ter-qPCR be validated on a cohort of clearly described LB patients and healthy controls from both North America and Europe before we could draw any conclusions on the diagnostic performance of the Ter-qPCR.