Vervolg op:
ICLB 2022 Amsterdam; Bron
viewtopic.php?f=5&t=2595#p28185 en vervolg op; Bron
viewtopic.php?f=5&t=2595#p28585 
In herinnering en
ter verificatie:
17 januari 2023:
Brochure Lymevereniging -
ICLB & Patiëntensymposium 2022; Bron
https://lymevereniging.nl/wp-content/up ... 221115.pdf..
Patiënten met moeilijk te diagnosticeren symptomen kunnen op zoek gaan naar alternatieven in Nederland en het buitenland:
-
Donkerveldmicroscopie-
Bioresonantie- Diverse PCR's (
Phelix Phage PCR/fagentest)
-
Cellulaire testen..
..
Onderzoeksresultaten - SubanalysesPraktische problemen:
cellulaire tests zijn foutgevoelig. De gevoeligheid na zes en twaalf weken is niet merkbaar verbeterd. De cellulaire tests presteren vergelijkbaar tussen:
- Patiënten met vroege lyme (erythema migrans) versus gedissemineerde lyme.
- Gezonde personen met een eerdere tekenbeet/Borrelia-antilichamen versus die zonder.
Combinatie van een antistoftest met een cellulaire test maakt deze niet beter.
Bij het testen van lymeziekte is in het algemeen de groep met gezonde personen (zonder lyme) veel groter dan de groep met
lymepatiënten. De onderzoekers stellen dat veel patiënten
vals-positieve testresultaten krijgen met de door hen onderzochte testen.
ConclusieDe onderzochte
cellulaire tests zijn ongeschikt voor gebruik,
vanwege:
-
lage gevoeligheid bij zowel vroege als gedissemineerde lyme.
-
lage specificiteit..
Interessante nieuwe
Publicatie!
Validatie van de
Phelix Phage Borrelia test van Red Laboratories (België)!
Elsevier ScienceDirect
Ticks and Tick-borne Diseases
Volume 16, Issue 4, July 2025, 102488; Bron
https://www.sciencedirect.com/journal/t ... 16/issue/4Publicatie Juli 2025Original article -
'The real-time PCR targeting the phage terminase (terL) is not suitable for diagnostics of human Borrelia infections in Europe' by Manja Zimmermann, Gabriele Margos, Christine Hartberger, Reto Lienhard, Anna J. Henningsson, Malin Lager, Mateusz Markowicz, Anna-Margarita Schötta, Andreas Sing, Benoit Jaulhac, Per-Eric Lindgren, Alje P. van Dam, Joppe W.R. Hovius, Volker Fingerle; Bron;
https://www.sciencedirect.com/science/a ... 9X25000524 en Bron;
https://pubmed.ncbi.nlm.nih.gov/40516432/ ..Highlights
- Evaluation of a real-time PCR targeting the large subunit of phage terminase (terL).
- terL PCR does not work equally well on all Borrelia species.
- not all isolates of B. afzelii and B. garinii are being detected.
- serum samples of patients with Lyme borreliosis were all PCR negative.
- terL PCR does not improve diagnostics of Lyme borreliosis patients in Europe..
..Abstract
Bacteria of the Borrelia burgdorferi sensu lato (sl) species complex can cause Lyme borreliosis (LB) in humans. PCR plays an important role in the diagnosis of many infectious diseases but it is used auxiliary in LB diagnostics. Here, we re-analysed a previously published real-time PCR targeting the multicopy gene of the large subunit of phage terminase (terL) in Borrelia. We analysed cultured material of Borrelia burgdorferi sl species, serum and clinical tissue samples of LB patients. PCR conditions were as previously described by Shan et al. 2021 but we also investigated PCR modifications.
PCR on cultured specimens showed that whilst all samples of B. burgdorferi sensu stricto (ss) gave a positive result, not all isolates of Borrelia species causing LB in Europe (i.e. B. afzelii, B. garinii) were detected by the terL PCR. Only slight differences in Ct values were detected between PCR runs using the original ZEN/IFBQ double quencher probe compared to other double quencher probes or single quencher probes. Contrary to the hypothesis expressed by the authors of the original paper that the PCR could detect phage DNA in serum, our data show that the terL PCR was negative on all tested serum samples of individuals diagnosed with proven LB. Furthermore, using patient’s tissue samples not all infections with B. afzelii or B. garinii were detected, similar to the results obtained with cultured material or serial DNA dilutions of Borrelia species. We conclude, that the terL PCR in its current form is unsuitable for LB diagnosis in Europe..
..
1. IntroductionPCR plays an important role in the diagnosis of many infectious diseases and it is used supportively in the diagnosis of Lyme borreliosis (LB), also known as Lyme disease. LB in humans is caused by bacteria of the Borrelia burgdorferi sensu lato (sl) species complex. PCR allows detection of DNA of these bacteria in patient samples and is useful to support or confirm other testing methods, such as antibody detection in blood, or in early disease before antibodies are detectable (Hofmann et al., 2017; Rauer et al., 2020). By detecting Borrelia DNA in tissue samples or body fluids, PCR can help to achieve an early diagnosis and treatment of LB. However, as there may be very few Borrelia bacteria in infected individuals and, hence, little DNA in tissue samples, a highly sensitive PCR is needed to provide reliable results.
In 2021,
Shan and co-authors (Shan et al., 2021) published a paper suggesting that a multicopy prophage gene, the large terminase subunit (terL), would improve diagnostics of LB when used as target in real-time PCR.
This PCR is being promoted for diagnostics of LB in humans (R.E.D. Laboratories Belgium, B-1731 Zellik, Belgium;
https://www.redlabs.com/p/home.html, access date 07 April 2025). Targeting multicopy genes for diagnostic purposes can be advantageous as it may increase the sensitivity of PCR and, therewith, improve diagnosis. In silico analyses suggested that the PCR may be suitable for all Borrelia species (Shan et al., 2021). However, in a letter to the editor the reproducibility of the statistical analyses and the specificity of the PCR were questioned (van de Schoor et al., 2021).
In a follow up paper (Shan et al., 2023), the usefulness of the approach was highlighted and expanded for detection of Borrelia
in ticks.
In
Europe, the most common Borrelia species that cause human LB are B. afzelii and B. garinii (Hofmann et al., 2017; Rauer et al., 2020), which are also the most prevalent Borrelia species found in ticks (Rauter and Hartung, 2005; Strnad et al., 2017). Therefore, a new PCR should detect those species.
In this study,
we have used the real-time PCR published by Shan et al. (
2021) in its original form and in a modified version to re-analyse clinical samples that were diagnosed as Borrelia positive in several laboratories using clinical assessment of the patients as well as p41 (Flagellin B (flaB)) (Schwaiger et al., 2001; Venczel et al., 2015) and outer surface protein A (ospA) (Ivacic et al., 2007; Briciu et al., 2016) as PCR targets. We also used cultured isolates of the Borrelia species commonly involved in clinical LB in Europe, i.e. B. afzelii, B. bavariensis, B. burgdorferi sensu stricto (ss), B. garinii, and B. spielmanii to analyse whether the PCR published by Shan et al. 2021 was suitable for detection of defined amounts of Borrelia DNA and to determine its detection limit. Published protocols for PCR on 16S rRNA (Gyllemark et al., 2021), flaB (Schwaiger et al., 2001), 5S-23S intergenic spacer (IGS) ((Strube et al., 2010), and unpublished) were used in parallel.
Analyses were performed independently by three different laboratories..
..2. Materials and methods
Samples included. DNA purified from in vitro cultures (n = 32) was included in the analyses. These included B. afzelii n = 10, B. burgdorferi ss (Bbss) n = 8, B. garinii n = 6, B. spielmanii n = 2, B. valaisiana n = 1, B. miyamotoi n = 1, B. recurrentis n = 2, B. hispanica n = 1, B. hermsii n = 1 (Table 1). We further analysed DNA of samples included in a panel that contained serial dilutions of DNA from nine Borrelia species (15 isolates) and single concentrations of specificity controls (four species, five isolates) (supplementary material Table S1). In Table 2 a subset of these samples is shown comparing the original terL PCR set up (master mix, double quencher probe) with a real-time PCR targeting a different locus (see Table heading for details). DNA dilutions ranged from 1 × 104 to 1 × 10–1 per 5 µl. Pre-treatment serum samples (Table 3a) from 15 patients with proven LB diagnosed by clinical assessment in combination with positive Borrelia-specific antibody titres or positive in vitro Borrelia culture from tissue biopsy were included. Furthermore, clinical samples (n = 26; Table 3b) of LB patients were included that had previously been diagnosed based on clinical symptoms, patient history and positive PCR using p41 (flaB) (Schwaiger et al., 2001; Venczel et al., 2015) and ospA (Ivacic et al., 2007; Briciu et al., 2016) as targets..
..4. Discussion
..In conclusion, use of the current form of the terL PCR assay cannot be recommended for the diagnosis of LB in patients in Europe and Asia. Furthermore, it is not suitable for detecting B. burgdorferi sl DNA in serum samples from infected individuals..
Conclusie & wake-up call: Veel mensen-patiënten zijn
jarenlang(
!) en worden tot op heden nog steeds
misleid!!!; Bron
viewtopic.php?f=38&t=2751#p30574 en; Bron
viewtopic.php?f=38&t=2751#p30595 en; Bron
viewtopic.php?f=38&t=2751&start=10#p30838 en; Bron
viewtopic.php?f=38&t=2751&start=20#p31137De ziekte van Lyme en chronische Lyme zijn een
verdienmodel geworden! En allerlei
commerciële partijen schromen er niet voor om over de rug van (erg) zieke mensen, Lyme patiënten of andere (zoekende, hoopvolle of wanhopige) zieke mensen veel geld te verdienen.
Patiënten moeten beter leren om zich zélf ook goed in de materie te leren verdiepen én
goed te leren lezen! En hetgeen dat wordt
gepromoot & geadviseerd door anderen (patiëntenorganisaties, patiënten, 'activistische Lyme organisaties&groepen', Ilads Lyme&me/cvs artsen, AVIG Lyme artsen, natuurartsen, therapeuten en behandelaars in privé-klinieken) goed te
verifiëren en daarmee het risico te vermijden om
ergens (
onwetend, in goed vertrouwen, naïef, 'blind')
achter aan te gaan lopen of achter zogenoemde op een voetstuk geplaatste
'Lyme-helden' aan te gaan lopen.
Conclusie: Met de kostbare
experimentele en niet-gevalideerde Phelix Phage Borrelia test van Red Laboratories (België) kan
géén diagnose Borrelia burgdorferi sensu lato (s.l) en of B. miyamotoi/Relapsing Fever worden gesteld!
De
gevolgen voor de (
geteste en te testen) patiënten zijn: een onjuiste testuitslag, een gemiste diagnose, een verkeerd gestelde diagnose, een verkeerde voorgeschreven behandeling, veroorzaakte (blijvende) schade aan de gezondheid.
Lees en leer meer informatie ter
verificatie over het
'het (onterecht!!) aanprijzen en promoten van de Phelix Phage Borreliat test van Red Laboratories (België) vanaf 2018 door de Lymevereniging, Stichting Lymefonds/het Lymefonds en sommige mensen uit de achterban'; Bron
viewtopic.php?f=38&t=2751&start=30#p31209 en Bron;
viewtopic.php?f=38&t=2751&start=30#p31210 Lees en leer meer en ter
verificatie: over
hoe de geschiedenis van het
'xmrv/mlv debacle' zich duidelijk heeft herhaald!; Bron
viewtopic.php?f=38&t=2751&start=70#p31612 en Bron;
viewtopic.php?f=37&t=2772&start=10#p30952 en Bron;
viewtopic.php?f=38&t=2435&start=140#p27534 En over
hoe er
veel geld wordt verdiend over de rug van (erg) zieke mensen!; Bron
viewtopic.php?f=38&t=2751&start=60#p31517Het
commerciële laboratorium Red Laboratories (België) is meerdere keren in opspraak geweest en het artikel van
Apache van
2017 (
klik op 'Lees alleen dit artikel gratis'); Bron
https://www.apache.be/2017/10/04/de-chr ... e-meirleir heeft duidelijkheid gegeven en is onderbouwd met officiële documenten.
Wie een goede
juridische¬ariële kennis heeft met name van het rechtsgebied
Ondernemingsrecht weet dat de (ondertekende) documenten als de oprichtingsakte, document consent of Directors bestuurdersfunctie van Directeur, het certificaat aandeelhouders, deelnemingsbewijs van R.E.D. Laboratories in REDLABS USA in functie van president, het vonnis van de rechtbank in de V.S Houston, Texas en het uittreksel van de Kamer van Koophandel
officiële juridische¬ariële documenten zijn waarvan de inhoud voor zich spreekt.
In herinnering:..
Leer en lees meer: ter
verificatie:
Frontiers in MicrobiologyOPINION article
Publicatie 13 december 2021Opinion:
'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/full en; Bron
https://pmc.ncbi.nlm.nih.gov/articles/PMC8710760/ en; Bron
viewtopic.php?f=5&t=2322&start=750#p27953..We 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..
..
ConclusionsWe 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..
Teulières (
natuurarts;
géén wetenschapper & onderzoeker!!) en de collega onderzoekers/wetenschappers van
Phelix Research & Development (Engeland); Bron
https://phelix.info/about/team/ &
Phelix France (Frankrijk); Bron
https://zb14927861.phelix.org.uk/team/ en; Bron
https://zb14927861.phelix.org.uk/about-us/ en Mijatovic (Red Laboratories België) hebben tot nu toe (nog) niet gereageerd op de Publicaties van het team wetenschappers-onderzoekers.
