Zijn er ervaringen met de Phelix Phage Borrelia test?

[Roxy]

Vervolg op; Bron viewtopic.php?f=38&t=2435&start=230#p30925 en vervolg op; 'Publicatie Validatie van de test'; Bron viewtopic.php?f=38&t=2435&start=230#p31504

Stand van zaken - European Patent Office (EPO) - WO2022072615 (EP21876463/EP4221827) 'Method of manipulating phages within the body to attack infectious target cells'; Bron https://register.epo.org/application?number=EP21876463 en; Bron https://register.epo.org/application?nu ... ab=doclist

Stand van zaken 5 november 2025: - European Patent Office (EPO); Bron https://www.epo.org/

..05.11.2025: New entry: Additional fee for renewal fee: despatch of communication + time limit..

Er moet €1.500 worden betaald. Zal de procedure worden voortgezet? Of zal de aanvrager Jernigan (natuurarts, géén onderzoeker & wetenschapper) van Biologix Center (Verenigde Staten) het dossier laten sluiten?

[Roxy]

Vervolg op: 'Interessante nieuwe Publicatie! Validatie van de test!'; Bron viewtopic.php?f=38&t=2435&start=230#p31504

Letters to the Editor
6 januari 2026
Article: 'Response to the critique on the TerL-based PCR by Zimmermann et al. (2025)' by Jinyu Shan, Tanja Mijatovic, Louis Teulières, Martha R.J. Clokie; Bron https://www.sciencedirect.com/science/a ... via%3Dihub

Letters to the Editor
6 januari 2026
Article: 'Response letter to Rebuttal to Zimmermann et al. (2025) critique of TerL-based PCR' by Manja Zimmermann, Gabriele Margos, Christine Hartberger, Reto Lienhard, Anna J. Henningsson, Malin Lager d, 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 ... via%3Dihub

..Dear Editor,
In response to the Rebuttal letter to our paper Zimmermann et al. (2025) titled “The real-time PCR targeting the Phage terminase (terL) is not suitable for diagnostics of human Borrelia infections in Europe”, we would first like to clarify that we do not and did not question the method of targeting the phage terminase large subunit gene (terL) by qPCR per se. We have critically evaluated the method on DNA extracted from the main European Borrelia species and found that it did not produce positive results for all isolates of B. afzelii and B. garinii. This evaluation is independent of DNA purification methods as control PCRs clearly showed that there was DNA of sufficient quality and quantity for PCR in these tests. It also clearly demonstrates that the terL PCR in its published design (Shan et al. 2021) is not suitable for detection of all Borrelia species and isolates. The follow-up publication of Shan et al. 2023 did not provide information on better design of primers and probes for B. burgdorferi sensu lato species, in fact it published primers and probes specific for Borrelia miyamotoi, a relapsing fever species vectored by Ixodes ticks. Further, it was a hypothesis put forward by Shan et al. 2021 that phages are released spontaneously into the bloodstream of infected individuals.

Below are our responses to the specific points that the authors raised:
1. Zimmermann et al. (2025) used the wrong sample type for terL PCR
The authors maintain that the wrong type of sample was used (serum instead of blood). However, we would like to emphasize that we did not use just one type of sample but several. We used:
1. DNA purified from cultured material of several Borrelia species known to occur in Europe
2. A DNA panel comprising of serial dilutions of DNA of several Borrelia species
3. DNA purified from tissue samples of patients diagnosed with Lyme borreliosis
4. Serum samples of patients diagnosed with Lyme borreliosis

In these samples we found that the real-time PCR designed by Shan et al. 2021 did not produce positive results for all B. afzelii or B. garinii samples tested. Control PCRs showed that the DNA was of sufficient quality and quantity and without PCR inhibiting substances to detect all Borrelia isolates. The serial dilution samples were tested in three laboratories independently with the same results. These results suggest a problem in the terL real-time PCR design and set up itself - independent from the sample type or extraction method.

In their original article (Shan et al. 2021) and in a follow-up review from 2024 (Teulières/Jia et al. 2024) the authors put forward that free phages circulate in the blood and can be detected in higher frequency in blood than in serum samples. However, in their supplementary table they described at least one reactive PCR in 6/13 early, 18/23 healthy and 35/42 late patients (without further defining “early” or “late” or what they would define as a positive result) suggesting that serum can also be used. In addition, in a paper published by Shan et al. 2024 in Methods in Molecular Biology, the authors themselves indicate that human serum can be used for DNA extraction: “Add 300 μL of human serum samples to suitable Eppendorf tubes.”

Furthermore, the proposal to use blood or serum for Borrelia diagnostic using the qPCR targeting the terL locus is based on the assumption that phages are spontaneously released and circulate in the blood stream or body fluids. However, this is a hypothesis that was supposedly demonstrated by an artificial in vitro setting (Shan et al. 2021). To the best of our knowledge, it has not yet been demonstrated that Borrelia phages are spontaneously induced and circulate free in the blood of infected individuals.

2. Zimmermann et al. (2025) used the wrong DNA extraction method for terL PCR
It may be so that phenol-chloroform extraction might work slightly better for DNA extraction from blood than the methods used in Zimmermann et al. 2025. However, in our evaluation we actually used DNA that was extracted from in vitro cultured bacteria. Quality and quantity of DNA and absence of PCR inhibitors was demonstrated using real-time PCRs that successfully amplified single copy loci of Borrelia. Thus, the failure of the real-time PCR targeting the terL locus to produce positive results for some B. afzelii and B. garinii isolates could not be due to using a possible wrong DNA extraction method, yet due to a problem inherent to the Shan et al. PCR design.

3. Zimmermann et al. fail to acknowledge proof-of-concept context and advances in phage-based methodologies
We do not fail to acknowledge the potential advantages of a phage-based or blood/serum-based diagnostic procedure for Borrelia in Lyme borreliosis patients and we are aware of potential shortcoming of some diagnostic procedures. However, with our critical evaluation of the terL qPCR we aimed to independently put a new diagnostic test under scientific scrutiny.

Contrary to the authors claim that the 2021 study was a preliminary study and that they “have developed new primers and probes targeting homologous phage genes in B. afzelii, B. garinii, and other genospecies common in Europe“, we did not find any further publications that would support this claim. In fact, a publication in Methods in Molecular Biology by Shan et al. in 2024 reports the same method, same primers, probes and procedures as the 2021 publication. If the study by Shan et al. 2021 and the method published by Shan et al. 2024 were preliminary, it should be clarified so that laboratories wanting to use the method are aware of it. From accessible public information it was not obvious to us that the test had been updated. Thus, “reviewing updated documentation” as suggested in the letter was not possible. None of the references given on the website of R.E.D. Laboratories (as per 25.09.2025: Shan et al. 2021, Targeting Multicopy Prophage gene for increased detection of Borrelia burgdorferi sensu lato (s.l.) the causative agents of Lyme borreliosis, in blood; Shan et al. 2023 Combining citizen science and molecular diagnostic methods to investigate the prevalence of Borrelia burgdorferi s.l. and Borrelia miyamotoi in tick pools across Great Britain; Jia et al. 2023 Overcoming the Challenge of Lyme disease diagnosis: The role of phage-based testing; Jia et al., 2024. Successful diagnosis and treatment of Borrelia miyamotoi in patients with joint and muscle pains, ME/CFS and cognitive dysfuntion following tick bites: a case report) mention a further development of specific primers and probes for B. afzelii or B. garinii or any other Borrelia burgdorferi s.l. species.

Of note, we did not investigate Borrelia miyamotoi or relapsing fever directed approaches in Zimmermann et al. 2025. This “broader diagnostic value” would need to be investigated in an additional study.

4. Zimmermann et al. (2025) failed to follow the scientific practice of faithful protocol reproduction
Any diagnostic PCR will need to be sufficiently robust to be performed in different laboratories. We tested the terL qPCR published by Shan et al. 2021 using the same primers, probes, master mixes in combination with DNA of various Borrelia species that did not contain inhibitors. We tested the PCR in three different laboratories that set up the PCR independently but used identical DNA serial dilutions for comparability. Each lab used their own control PCR and came to the same conclusion: the terL PCR published by Shan et al. 2021 did not give positive results with some isolates of B. afzelii and B. garinii. Thus, again we can only conclude that our data suggest that the lack of sensitivity is inherent in the qPCR protocol published by Shan et al. 2021, 2024.

5. Conclusion: collaboration, rigorous replication, and further development
We have published the results of our scientific evaluation of the real-time PCR targeting the terL locus so that differences or weaknesses can be acknowledged and the scientific and medical community is aware of it. We very much welcome research into improved diagnostic tests for Lyme borreliosis and independent validation of such tests in other settings and an open, transparent and professional discussion about these data are crucially important.

Finally, we would also like to point out a commentary that addresses other methodological deficiencies in the study by Shan et al. 2021 (Van de Schoor et al. 2021; Bron https://www.frontiersin.org/journals/mi ... 02131/full )..

[Roxy]

Vervolg op; Bron viewtopic.php?f=38&t=2435&p=32499#p32167 en vervolg op: Publicatie Validatie van de test; Bron viewtopic.php?f=38&t=2435&start=230#p31504 en vervolg op: Letters to the Editor; Bron viewtopic.php?f=38&t=2435&p=32790#p32499

Stand van zaken - European Patent Office (EPO) - WO2022072615 (EP21876463/EP4221827) 'Method of manipulating phages within the body to attack infectious target cells'; Bron https://register.epo.org/application?number=EP21876463 en; Bron https://register.epo.org/application?nu ... ab=doclist

Stand van zaken 22 april 2026: - European Patent Office (EPO); Bron https://www.epo.org/

..22.04.2026: New entry: Application deemed to be withdrawn: despatch of communication + time limit..

De aanvraag van Jernigan (natuurarts, géén onderzoeker & wetenschapper) van Biologix Center (Verenigde Staten) wordt geacht te zijn ingetrokken (niet-betaling van de verlengingskosten). (..en in herinnering: 'Phelix Borrelia-Phage Test: A True Game-Changer in the Fight Against Lyme Disease'; Bron https://biologixcenter.com/lab-testing/ ... e-disease/ en: 'Not Every Lyme Disease Lab Test Is Very Sensitive - Why the Phelix Borrelia Phage Test is The New Gold Standard for Lyme Disease and Relpasing Fever'; Bron https://biologixcenter.com/inpt-phage-t ... r-testing/ en: de 'EM signature' en de 'BES findings'; Bron viewtopic.php?f=38&t=2435&start=220#p30682 ..).

Over Conferenties en Webinars van Ilads of van CIP of van AONM of van Lyme Resource Centre (LRC).

En over hoe de behandelaars (behandelaars in privé-klinieken, Ilads- en AVIG Lyme artsen, natuurartsen en therapeuten) én de patiënten (wereldwijd) tot op heden nog steeds worden misleid!!!

2026: Ilads European Scientific Conference London (UK), 16-18 April 2026; Bron https://membercompass.ilads.org/events/ ... conference De Sprekerslijst; Bron https://membercompass.ilads.org/ilads-c ... e-speakers En de Agenda; Bron https://assets.noviams.com/novi-file-up ... -26-26.pdf

..16 April 2026: 4:00-4:45 PM - Diagnostics - Tanja Mijatovic, PhD..

En over hoe er veel geld wordt verdiend over de rug van (erg) zieke mensen!; Bron viewtopic.php?f=38&t=2751&start=60#p31517 De geschiedenis van het 'xmrv/mlv debacle' heeft zich duidelijk herhaald!; Bron viewtopic.php?f=37&t=2772&start=10#p30952 en Bron; viewtopic.php?f=38&t=2435&start=140#p27534

Het 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&notariële 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&notariële documenten zijn waarvan de inhoud voor zich spreekt.

*Red Laboratories (België) is een commercieel laboratorium en heeft géén ISO 15189 accreditatie van het BELAC België (de Belgische Accreditatie-instelling - Medische laboratoria (MED); Bron https://economie.fgov.be/nl/themas/kwal ... atoria-med

Ter verificatie:
ILAC Mutual Recognition Arrangement (ILAC MRA); Bron https://ilac.org/ en de geregistreerde leden ILAC MRA Signatory Search; Bron https://ilac.org/signatory-search/
- BELAC België (Belgian Accreditation Structure) is lid van ILAC MRA; Bron https://ilac.org/signatory-detail/?id=8 maar Red Laboratories - België heeft géén ISO 15189 accreditatie van het BELAC België (Belgian Accreditation Structure).
Lees en leer meer; Bron viewtopic.php?f=38&t=2751#p30583 en; Bron viewtopic.php?f=38&t=2751#p30594 en; Bron viewtopic.php?f=38&t=2435&start=220#p30236 en; Bron viewtopic.php?f=38&t=2435&start=220#p30237

[Roxy]

Vervolg op; Bron viewtopic.php?f=38&t=2435&start=240#p32790 en vervolg op: Publicatie Validatie van de test; Bron viewtopic.php?f=38&t=2435&start=230#p31504 en vervolg op: Letters to the Editor; Bron viewtopic.php?f=38&t=2435&p=32790#p32499

Stand van zaken - European Patent Office (EPO) - WO2022072615 (EP21876463/EP4221827) 'Method of manipulating phages within the body to attack infectious target cells'; Bron https://register.epo.org/application?number=EP21876463 en; Bron https://register.epo.org/application?nu ... ab=doclist

Stand van zaken 17 juli 2026: - European Patent Office (EPO); Bron https://www.epo.org/

..17.07.2026: New entry: Application deemed to be withdrawn - published on 19.08.2026 [2026/34]..
..21.07.2026: Refund of fees..

De aanvraag van Jernigan (natuurarts, géén onderzoeker & wetenschapper) van Biologix Center (Verenigde Staten) is ingetrokken en dat wordt door het European Patent Office (EPO) op 19 augustus 2026 gepubliceerd in de Editie [2026/34].

Stichting Tekenbeetziekten - De diagnose van de ziekte van Lyme; Bron https://www.tekenbeetziekten.nl/de-teek ... /diagnose/

..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. Meer gedegen wetenschappelijk onderzoek is dus gewenst! We volgen de ontwikkelingen met belangstelling..

..'de test is niet gevalideerd'..?

Het gestelde is onjuist! De test is gevalideerd! en is onbetrouwbaar gebleken! Conclusie: Met de kostbare test 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.

Leerzaam: en ter verificatie: Validatie van de test Publicatie Juli 2025
Original 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..

En leerzaam: en ter verificatie: Letters to the Editor; Bron viewtopic.php?f=38&t=2435&p=32790#p32499

Het is van groot belang dat de Lyme patiëntenorganisaties in Nederland de (nieuwe) Lyme patiënten of chronische Lyme patiënten of andere (zoekende, hoopvolle of wanhopige) zieke mensen in Nederland duidelijk & goed informeren en de juiste informaties verstrekken! En dat de mensen-patiënten niet tot op heden nog steeds worden misleid!

En in herinnering: 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

En in herinnering: de geschiedenis van het 'xmrv/mlv debacle' heeft zich duidelijk herhaald!; 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 (érg) zieke mensen!; Bron viewtopic.php?f=38&t=2751&start=60#p31517
En in herinnering: lees en leer meer: over de misleiding door de jaren heen; Bron viewtopic.php?f=6&t=3475&p=32888#p32845

Conclusie & wake-up call: Er wordt helaas véél onjuiste informatie verspreid op internet, fora, in blogs of in Lymegroepen. En daarom moeten patiënten beter leren om zich zélf ook goed in de materie te leren verdiepen én goed te leren lezen!!! ( de informaties, de medische artikelen, de Proefschriften en ook de Publicaties van de Resultaten van Wetenschappelijk onderzoek). 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' (of zogenoemde 'Lyme-helden' die géén 'Lyme-helden' blijken te zijn!!) aan te gaan lopen.

Lees en leer meer: Wat is wetenschappelijk onderzoek en waarom is het nodig? Hoe werkt wetenschappelijk onderzoek?; Bron viewtopic.php?f=6&t=3485&p=32915#p32908