Symposium voor wetenschappers en medici; Bron viewtopic.php?f=5&t=2595#p28185
Programma; Bron https://congresscare.com/congress/16th- ... iseases-2/
Topics; Bron https://www.iclb2022.org/program-topics/
Congress committees; Bron https://www.iclb2022.org/committees/
BMC Microbiology volume 23, Article number: 204 (2023)
Publicatie 1 augustus 2023
Open access - 'Lyme borreliosis diagnosis: state of the art of improvements and innovations' by Mickaël Guérin, Marc Shawky, Ahed Zedan, Stéphane Octave, Bérangère Avalle, Irene Maffucci & Séverine Padiolleau-Lefèvre; Bron https://bmcmicrobiol.biomedcentral.com/ ... 23-02935-5
..Abstract
With almost 700 000 estimated cases each year in the United States and Europe, Lyme borreliosis (LB), also called Lyme disease, is the most common tick-borne illness in the world. Transmitted by ticks of the genus Ixodes and caused by bacteria Borrelia burgdorferi sensu lato, LB occurs with various symptoms, such as erythema migrans, which is characteristic, whereas others involve blurred clinical features such as fatigue, headaches, arthralgia, and myalgia. The diagnosis of Lyme borreliosis, based on a standard two-tiered serology, is the subject of many debates and controversies, since it relies on an indirect approach which suffers from a low sensitivity depending on the stage of the disease. Above all, early detection of the disease raises some issues. Inappropriate diagnosis of Lyme borreliosis leads to therapeutic wandering, inducing potential chronic infection with a strong antibody response that fails to clear the infection. Early and proper detection of Lyme disease is essential to propose an adequate treatment to patients and avoid the persistence of the pathogen. This review presents the available tests, with an emphasis on the improvements of the current diagnosis, the innovative methods and ideas which, ultimately, will allow more precise detection of LB..
..Traditional PCR assays and PCR improvements
Moreover, bacteriophages have a specific tropism towards certain bacteria. In other words, phages infect a specific bacterial species and their genetic material is specific to the bacteria they infect, resulting in a high specificity of the test exploiting this approach, **such as the Phelix Phage test patented by Dr. Louis Teulières [196].
Even if this technique is promising, **an analysis of this study shows inaccuracies in terms of statistical analysis or cohort composition and comparison with healthy controls, thus requiring validation [199]..
..ELISpot and derived approaches
Many studies show that Borrelia burgdorferi can enter endothelial cells and macrophages [90,91,92]. This ability is one of the main components of the humoral immune escape mechanism of Bb [27]. Similarly to viral infections, intracellular activation of type I interferons (IFN) plays an important role in B. burgdorferi infection [93]. One of the immune responses of the host in LB infection is characterized by a cytokine response and the secretion of IFN-γ followed by a high expression of IL-4, by T helper 1 lymphocytes, which are associated with non-chronic manifestations of LB. However, persistent IFN-γ expression could lead to chronicity of the LB [94, 95].
In light of this, the secretion of IFN-γ can be exploited to detect the presence of Ag from Bb using ELISpot methods [96]. It measures the antigen-specific cellular responses by quantifying the number of IFN-γ-producing T cells [97]. The sensitivity of this kind of method is known to be about 20 to 200 times greater than ELISA or flow cytometry, and it is based on conditions (antigens and cellular medium) used for LT activation [98]. The iSpot Lyme™ is an ELISpot method ***(Autoimmun Diagnostika/Genome Identification Diagnostics) that uses recombinant Borrelia antigens (recombinant DbpA, OspC, p100, and VlsE) to stimulate specific effector/memory T cells combined to a signal enhancer medium CTL Test Plus. This assay showed significantly higher sensitivity (84%) as compared to the WB (30%) [76]. However, as for conventional serology, traditional or improved ELISPOT assays cannot differentiate active from past LB ***[99].
The ELISpot principle has been combined with the QuantiFERON technology (QIAGEN Sciences) for the detection and measurement of IFN-γ in the case of Mycobacterium tuberculosis infection [100]. Taking inspiration from this technology, Callister’s team developed the QuantiFERON-Lyme assay, which consists in the detection of IFN-γ in whole blood after incubation with synthetic Borrelia antigens (p66, DbpB, OspC, and flagellin) [78]. This test showed a sensitivity of approximately 70% in patients with EM versus 17% for standard serology. In addition, this test allows the distinction between an active or past infection because of the significant decrease of the immune response in patients who have been treated. However, this technique seems to be suitable only in the United States where Bbss, one of the species belonging to the B. burgdorferi sensu lato complex, dominates. In Europe, where B. afzelii and B. garinii are the dominant species, induced responses are low and a more sensitive test is therefore required [101]. The use of INF-γ for diagnosis could be challenging, and dependent on the cell type used for the assay. Indeed, a recent study confirmed that Borrelia burgdorferi is a poor inducer of INF-γ production by peripheral blood mononuclear cells (PBMCs) [102] but a strong inducer by human primary NK cells [103]. More studies have to be done regarding specificity and reliability.
Many other methods are based on the ELISpot principle, such as the Spirofind Revised (Oxford Immunotec), which quantifies the IL-1ß produced by primary PBMC cells after contact with Borrelia mix, using bead ELISA assay [104].
Also based on T cells, some authors exploited the massive sequencing of T-cell receptor repertoire to highlight the specificity of T-cell responses and probe pathogen exposure. Indeed, it is known that the serological approach based on specific antibody-response suffers from a seronegative window period of 2 to 4 weeks [105], whereas T-cell response is detectable before the humoral response [79]. To circumvent both the co-infection issue and the difference between the kinetics of T-cell response and the humoral response, Greissl and collaborators recently proposed an aid for diagnosis based on a tool allowing to analyze and classify TCR sequencing in order to ensure higher sensitivity testing compared with STTT, in particular concerning early LB (Table 3).
Other tests also based on cellular proliferation are available. A few years ago, a Lymphocyte Transformation Test-Memory Lymphocyte Immunostimulation Assay (LTT-MELISA®, InVitaLab) was developed. This assay is based on two steps: uptake of radioisotope by dividing lymphocytes, followed by their detection [106]. This method evaluates the lymphoproliferative response of PBMCs to B. burgdorferi antigens. iSpotLyme™, Spirofind™, and LTT-MELISA® assays have been recently studied and have been compared to classical serology testing [104]. ****According to the sensitivity and specificity reported in Table 3, it has been confirmed that these cellular tests lead to a high number of false-positive results and are unfit for clinical use at this stage ****[77]..
**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/full en Bron viewtopic.php?f=5&t=2322&start=750#p27953
..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..
***8 december 2022 Promotie (Universiteit Leiden); Bron https://www.universiteitleiden.nl/agend ... orreliosis 'Proefschrift Gorkom, T. van (2022): Immunodiagnostics of Lyme neuroborreliosis'; Bron https://scholarlypublications.universit ... 87/3494393
Clinical and Experimental Immunology
Publicatie 7 januari 2020
Journal Article - 'Prospective comparison of two enzyme-linked immunosorbent spot assays for the diagnosis of Lyme neuroborreliosis' by T van Gorkom, W Voet, S U C Sankatsing, C D M Nijhuis, E ter Haak, K Kremer, S F T Thijsen; Bron https://academic.oup.com/cei/article/199/3/337/6402722
Journal of Clinical Microbiology..The results of a comparative pilot experiment that we performed in which we assessed the influence of the deviations discussed above supported that these deviations from the recommended protocol are not critical as such (Supporting information, Data S4). Hence, the conclusion stands that both ELISpot assays cannot help to diagnose active LNB..
Publicatie 24 januari 2018
Prospective study - 'An Enzyme-Linked Immunosorbent Spot Assay Measuring Borrelia burgdorferi B31-Specific Interferon Gamma-Secreting T Cells Cannot Discriminate Active Lyme Neuroborreliosis from Past Lyme Borreliosis: a Prospective Study in the Netherlands' by T. van Gorkom, S.U.C. Sankatsing, W. Voet, D.M. Ismail, R.H. Muilwijk, M. Salomons, B.J.M. Vlaminckx, A.W.J. Bossink, D.W. Notermans, J.J.M. Bouwman, K. Kremer, S.F.T. Thijsen; Bron https://journals.asm.org/doi/pdf/10.1128/JCM.01695-17
TVI clinic Tijdschrift voor Infectieziekten..In conclusion, the Borrelia ELISpot assay used in this study, measuring the number of B. burgdorferi B31-specific IFN-secreting T cells, cannot be used for the diagnosis of active Lyme neuroborreliosis..
Jaargang 14, Nummer 3, Juni 2019; Bron https://www.tvionline.nl/journal-editio ... juni-2019/
Publicatie - 'Geen onderscheid tussen actieve Lyme-neuroborreliose en Lyme-borreliose uit het verleden met cellulaire ELISpot-test'; Bron https://www.ariez.nl/wp-content/uploads ... Gorkom.pdf
..SAMENVATTING
De diagnostiek van Lyme-borreliose (LB) is gebaseerd op het aantonen van Borrelia-specifieke anti-stoffen, maar een positieve testuitslag is geen bewijs voor actieve ziekte. Om actieve LB te kunnen vaststellen is betere diagnostiek nodig. Recentelijk zijn testen ontwikkeld die de activiteit van het cellulaire immuunsysteem meten, maar de klinische validatie van dergelijke testen bij goed gedefinieerde patiëntengroepen ontbreekt. Deze prospectieve studie beschrijft de validatie van een ‘enzyme-linked immunosorbent spot’- (ELISpot-) test voor de diagnose van een actieve Lyme-neuroborreliose. De studie toont aan dat de Borrelia-ELISpot-test correleert met blootstelling aan de Borrelia-bacterie, maar niet kan worden gebruikt voor de diagnose van een actieve Lyme-neuroborreliose..
****The Lancet Infectious Diseases - Online First; Bron https://www.thelancet.com/journals/laninf/onlinefirst
Publicatie 14 juni 2022
Article - 'Diagnostic parameters of cellular tests for Lyme borreliosis in Europe (VICTORY study): a case-control study' by E Baarsma, MD, Freek R van de Schoor, MD, Stefanie A Gauw, RN, Hedwig D Vrijmoeth, MD, Jeanine Ursinus, MD, Nienke Goudriaan, MD, Calin D Popa, MD, Hadewych JM ter Hofstede, MD, Mariska MG Leeflang, PhD, Kristin Kremer, PhD, Cees C van den Wijngaard, PhD, Prof Bart-Jan Kullberg, MD, Prof Leo AB Joosten, PhD, Prof Joppe W Hovius, MD
En; Bron viewtopic.php?f=5&t=2595&start=190#p29459 en; Bron viewtopic.php?f=5&t=2595&start=210#p29525 en; Bron viewtopic.php?f=5&t=2322&start=1030#p29463 En aanvullend op 'Stand van zaken Phelix Phage Borrelia test'; Bron viewtopic.php?f=38&t=2435&start=190#p29587..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..
Ook Innatoss Laboratories berichtte in 2018 over het gedane validatie onderzoek door Innatoss met de EliSpot/LymeSpot test (Autoimmun Diagnostika GmbH) en over het verrichte validatie onderzoek.
Innatoss Laboratories is het helaas ook niet gelukt om een goede nieuwe cellulaire test te kunnen ontwikkelen en is gebleken uit de resultaten van het validatie onderzoek bij de Ixodus-studie (Ixodus test); Bron https://www.innatoss.com/nl/onderzoek/studie-ixodes. Dat getuigt van een goede informatieverstrekking aan de mensen-patiënten en de artsen. En getuigt van een professionele werkmethode waar andere commerciële laboratoria nog een voorbeeld aan kunnen nemen.Bron viewtopic.php?f=38&t=1799&start=130 Innatoss 4 december 2018: 'We have used LymeSpot. Fortunately, no 80% positives. This would make a test useless. However LymeSpot does not differentiate between active and past infection. Tamara van Gorkom found the same. VICTORY study is ongoing in Amsterdam; Bron https://twitter.com/InnatOss