bioRxiv - Pathology最新文献

{"title":"The amyloid oligomer modulator anle138b has disease modifying effects in a human IAPP transgenic mouse model of type 2 diabetes mellitus (hIAPP Ob/Ob mice)","authors":"Jo Hoppener, Christian Griesinger, Mohammed M.H. Alabariqi, Neville Vassallo, Niels Eijkelkamp, Armin Giese, Sanne M.G. Baauw, Sjors Fens, Sergey Ryazanov, Andrei Leonov, Hanneke L.D.M. Willemen, Bram Gerritsen, Nikolas Stathonikos, Lucie Khemtemourian, Raina Marie Seychell, Adam El Saghir, Sabine Versteeg","doi":"10.1101/2024.08.27.609850","DOIUrl":"https://doi.org/10.1101/2024.08.27.609850","url":null,"abstract":"Cytotoxic aggregates of human islet amyloid polypeptide (hIAPP) contribute to type 2 diabetes mellitus (T2DM) pathogenesis by damaging pancreatic islet β cells and reducing insulin production. Anle138b is an amyloid oligomer modulator with disease modifying properties in mouse models of neurodegenerative diseases linked to protein aggregation and with favorable results in phase 1 clinical studies. We tested whether anle138b has disease modifying properties in a severe hIAPP transgenic mouse model of T2DM. Oral administration of anle138b in hIAPP Ob/Ob mice reduced hyperglycemia, decreased glycated hemoglobin levels, increased islet β-cell mass and improved islet function compared to non-treated mice. In contrast, anle138b administration did not affect these parameters in non-transgenic Ob/Ob mice, indicating that the anti-diabetic effects of anle138b are hIAPP-dependent. In vitro, anle138b inhibited hIAPP aggregation and toxic effects of hIAPP on mitochondria. These results indicate that anle138b is a promising drug candidate for treating and/or preventing T2DM -associated pathology.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"33 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142183044","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Autosomal dominant Retinitis Pigmentosa caused by the rhodopsin isoleucine 255 deletion features rapid neuroretinal degeneration, decreased synaptic connectivity, and neuroinflammation.","authors":"Bowen Cao, Yu Zhu, Alexander Gunter, Ellen Kilger, Sylvia Bolz, Christine Hennes, Regine Muhlfriedel, Francois Paquet-Durand, Blanca Arango-Gonzalez, Marius Ueffing","doi":"10.1101/2024.08.29.610258","DOIUrl":"https://doi.org/10.1101/2024.08.29.610258","url":null,"abstract":"Retinitis Pigmentosa (RP) is a group of inherited retinal diseases that initially affects rod photoreceptors and causes progressive vision loss and blindness. Mutations in rhodopsin (RHO) can cause both autosomal recessive (ar) and dominant (ad) forms of RP, yet, the underlying degenerative mechanisms remain largely unknown, rendering the disease untreatable. Here, we focus on an in-frame, 3-base pair deletion, eliminating the isoleucine residue at codon 255 (i.e., RHOdeltaI255) and resulting in adRP. We generated a novel knock-in mouse homologous to the human RHOdeltaI255 mutation. This new mouse model displays a severe disruption of photoreceptor structure and function, as is seen in human patients. Our results indicate that this form of RP is a systems disease of the neuroretina that also impacts neuronal connectivity of bipolar- and horizontal cells, initiates neuroinflammation, and reduces the structural and functional integrity of the retina. Typical for adRP, RhodeltaI255 mice exhibit primary rod photoreceptor loss, followed by secondary cone degeneration, rhodopsin protein (RHO) mislocalization, progressive shortening of outer segments (OS), and disorganized OS structures. Subsequently, increasing gliosis, morphologic abnormalities of the inner retina, and impaired cone-driven visual function developed. In adRP, a single mutated allele is sufficient to cause the disease, as confirmed here in RhodeltaI255/+ heterozygous animals, where most photoreceptors were lost within two months after birth. Compared to this, homozygous RhodeltaI255/deltaI255 mutants exhibit an accelerated onset and even faster progression of retinal degeneration. The degeneration of RhodeltaI255-mutant photoreceptors was linked to the activation of both caspase- and calpain-type proteases, as well as poly(ADP-ribose) polymerase (PARP), indicating a parallel execution of both apoptotic and non-apoptotic processes. In conclusion, our data indicate that this form of RP affects the neuroretina beyond photoreceptor cell loss sharing features typical for other degenerative central nervous systems diseases, an insight, which may bear critical impact to understand and eventually develop treatment for these currently untreatable forms of blindness.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"124 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142183042","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multi-omics analysis of virus-permissive versus hostile cellular states reveals protein networks controlling virus infection","authors":"Honglin Chen, Philip D Charles, Quan Gu, Sabrina Liberatori, David L Robertson, Massimo Palmarini, Sam J Wilson, Shabaz Mohammed, Alfredo Castello","doi":"10.1101/2024.08.29.610331","DOIUrl":"https://doi.org/10.1101/2024.08.29.610331","url":null,"abstract":"The capacity of host cells to sustain or restrict virus infection is influenced by their proteome. Understanding the compendium of proteins defining cellular permissiveness is key to many questions in fundamental virology. Here, we apply a multiomic approach to determine the proteins that are associated with highly permissive, intermediate, and hostile cellular states. We observed two groups of differentially regulated genes: i) with robust changes in mRNA and protein levels, and ii) with protein/RNA discordances. Many of the latter are classified as interferon stimulated genes (ISGs) but have no reported antiviral activity. This suggests that IFN-dependent changes in mRNA levels do not imply antiviral function. Phosphoproteomics revealed an additional regulatory layer involving non-signalling proteins with altered phosphorylation. Indeed, we confirmed that several permissiveness-associated proteins with changes in abundance or phosphorylation regulate infection fitness. Altogether, our study provides a comprehensive and systematic map of the cellular alterations driving virus susceptibility.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"173 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142183094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Altered functional connectivity and spatiotemporal dynamics in individuals with sleep disorders","authors":"Lauren Daley, Prabhjyot Saini, Harrison Watters, Yasmine Bassil, Eric Schumacher, Lynn Marie Trotti, Shella Keilholz","doi":"10.1101/2024.08.28.610147","DOIUrl":"https://doi.org/10.1101/2024.08.28.610147","url":null,"abstract":"Idiopathic hypersomnia (IH) is a sleep disorder characterized by highly disruptive symptoms. Like narcolepsy type 1, a well-characterized sleep disorder, individuals with IH suffer from excessive daytime sleepiness, though there is little overlap in metabolic or neural biomarkers across these two disorders. This lack of common pathophysiology, combined with the clear overlap in symptoms presents an ideal paradigm for better understanding the impact of IH on an individual's functional activity and organization, and potentially, the underlying pathophysiology. This study examines the observed functional connectivity in patients with IH, and patients with narcolepsy type 1 (NT1) against healthy control individuals. Static functional connectivity is compared, as are quasi-periodic patterns, acquired from the BOLD timecourse, for all groups. In addition to baseline data comparison, the study also included a post-nap condition, where the individuals included in this analysis napped for at least 10 minutes prior to the scanning session, to explore why individuals with IH do not feel refreshed after a nap like individuals with NT1 do. Assessing the groups' spatiotemporal patterns revealed key differences across both disorders and conditions: static connectivity revealed at baseline higher subcortical connectivity in the NT1 group. There was also observably less connectivity in the IH group both at baseline and post-nap, though none of these static analyses survived multiple comparisons correction to reach significance. The QPP results however found significant differences in the IH group in key networks, particularly the DAN/FPCN correlation is significantly different at baseline vs. post-nap, a trend not observed in either the control or NT1 groups. The DAN and FPCN are drastically altered both at baseline and post-nap when compared to the other groups, and may likely be a disorder-specific result. This study demonstrates that key networks for arousal are more heavily disrupted in IH patients, who are less affected by a nap, confirmed through both subject reporting and functional evidence through spatiotemporal patterns.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"57 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142183092","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sex specific knee joint soft tissue mineralization with Fibrillin-1 mutation in male Tight Skin mice.","authors":"Craig Keenan, Xi Wang, Tayfun Dikmen, Yan Wen, Lorenzo Ramos-Mucci, Emily Shorter, David Abraham, George Bou-Gharios, Blandine Poulet","doi":"10.1101/2024.08.28.610095","DOIUrl":"https://doi.org/10.1101/2024.08.28.610095","url":null,"abstract":"Articular soft tissue mineralization and ossification are clear pathological signs of osteoarthritis joints. However their molecular and cellular aetiologies remain largely unknown. TGF-β family members are known contributors to both pathological ossification and osteoarthritis development. In this study, we used a Fibrillin-1 mutant mouse, the Tight Skin mouse (TSK), to define the detrimental effects of abnormal Fbn1 in TSK mice and known high TGF-β activity in joint pathology such as articular soft tissue mineralization and ossification. Knee joints of male and Female TSK and Wild-type (WT) littermates were analysed my micro-CT imaging and histology for articular soft tissue pathologies, as well as OA severity. Both aged (10, 26, 35 and 52wks) and following in vivo non-invasive repetitive joint overloading were used. We find that male TSK mice develop spontaneous soft tissue ossification from 26wks of age, followed by increased osteoarthritis at 1 year-old. In addition, knee joint overloading induced ligament and meniscal mineralisation and ossification in both WT and TSK male mice, but were significantly more severe in TSK knees, including ossification of the patella ligament and synovial lining. In contrast, female TSK knees did not develop more severe soft tissue mineralisation compared to littermate WT mice in neither aged nor overloaded knees. We conclude that Fbn1 mutation, and possibly overactive TGF-β activity in TSK mice, induce articular soft tissue ossification and osteoarthritis in a sex-specific manner. Further studies are needed to confirm the specific signalling involved and the relative protection from female mice from such pathologies.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"14 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142183090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Haemolymphatic tissues of captive boa constrictor (Boa constrictor): morphological features in healthy individuals and with Boid Inclusion Body Disease","authors":"Eva Dervas, Eleni Michalopoulou, Tanja Thiele, Francesca Baggio, Udo Hetzel, Anja Kipar","doi":"10.1101/2024.08.26.609690","DOIUrl":"https://doi.org/10.1101/2024.08.26.609690","url":null,"abstract":"Knowledge on the structure and composition of the haematopoietic tissue (HT) is essential to understand the basic immune functions of the immune system in any species. For reptiles, it is extremely limited, hence we undertook an in-depth in situ investigation of the HT (bone marrow, thymus, spleen, lymphatic tissue of the alimentary tract) in the common boa (Boa constrictor). We also assessed age- and disease-related changes, with a special focus on Boid Inclusion Body Disease , a highly relevant reptarenavirus-associated disease in boid snakes. The HT was subjected to gross, histological and ultrastructural examination, including special stains, immunohistochemistry, in situ hybridization and morphometric analyses. In general, the HT was dominated by T cells and lacked a clear structural organization, such as follicle formation. BIBD was associated with significantly higher cellularity and a granulomatous response in the spleen, and the presence of virus-infected haematopoietic cells in the bone marrow, suggesting the latter as a persistent source of viremia.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"131 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142183093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Vapor-based Fixation of Pulmonary Tissue in its Physiological State: A Novel Approach to Histological Validation of Ultra High Resolution Phase Contrast CT in Human Sized Lungs","authors":"Christian Dullin, Johanna Reiser, Willi Wagner, Elena Longo, Marko Prasek, Adriano Contillo, Nicola Sodini, Diego Dreossi, Paola Confalonieri, Francesco Salton, Marco Confalonieri, Elisa Baratella, Maria Cova, Claudia Benke, Md Motiur Rahman Sagar, Lorenzo D'Amico, Jonas Albers, Angelika Svetlove, Tatiana Flisikowska, Krzysztof Flisikowski, Mark O. Wielpuetz, Juergen Biederer, Hans-Ulrich Kauczor, Fabrizio Zanconati, Giuliana Tromba","doi":"10.1101/2024.08.26.609748","DOIUrl":"https://doi.org/10.1101/2024.08.26.609748","url":null,"abstract":"Lung diseases continue to present a major burden on public health. Therefore, improving the process of diagnosis by the development of novel imaging techniques is of great importance. In this perspective, phase sensitive CT imaging techniques such as propagation based imaging (PBI) might play an important role as they allow increasing the spatial resolution at very low x-ray dose levels that are comparable to clinical CT. However, the development of such methods is not only hindered by technological problems but also by the lack of precise validation strategies.\u0000We adapted formaldehyde (FA) vapor fixation to demonstrate that fresh porcine lungs that have been investigated by PBI can be fixed in their physiological shape and studied by multi-scale microCT imaging as well as classical histology. In addition, we show that FA vapor fixed pig lungs can be scanned by PBI without visible deterioration of image quality compared to fresh tissue. This opens the possibility of fixing and storing, for instance, human lung tissue before performing a PBI experiment, which in turn allows to study pathological changes in human lungs without questioning the translate-ability of findings in pig lung. The setup can be used by any interested researchers.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"49 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142183091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The p.H222P lamin A/C mutation induces heart failure via impaired mitochondrial calcium uptake in human cardiac laminopathy","authors":"Magali Seguret, Charlene Jouve, Andrea Ruiz-velasco, Lucille Deshayes, Zoheir Guesmia, Celine Pereira, Karim Wahbi, Jeremy Fauconnier, Gisele Bonne, Antoine Muchir, Jean-Sebastien Hulot","doi":"10.1101/2024.08.21.609073","DOIUrl":"https://doi.org/10.1101/2024.08.21.609073","url":null,"abstract":"Background: Mutations in the LMNA gene, which encodes lamin A/C, cause a variety of diseases known as laminopathies. Some mutations are particularly associated with the occurrence of dilated cardiomyopathy and heart failure, but the genotype-phenotype relationship and underlying mechanisms are unclear. Here, we used induced pluripotent stem cells (hiPSCs) from a patient carrying a LMNA point mutation (c.665A>C, p.His222Pro) to investigate the mechanisms leading to contractile dysfunction.\u0000Methods: LMNA p.H222P mutant and a CRISPR/Cas9 corrected isogenic control hiPSCs clones were differentiated into cardiomyocytes (hiPSC-CMs). Immunofluorescence staining was performed on hiPSC-CMs to quantify their sarcomere organization (SarcOrgScore) using a Matlab code. Ring-shaped cardiac 3D organoids were generated to compare the contractile properties of the two clones. Calcium transients in mutant and corrected hiPSC-CMs were measured by live confocal imaging. Mitochondrial respiration parameters were measured by Seahorse. Results: hiPSC-CMs were generated from the LMNA mutant and the corrected hiPSCs with no difference in the differentiation yield (proportion of troponin-positive cells: 95.0% for LMNA p.H222P vs. 95.1% for Ctrl-iso1, p=0.726). hiPSC-CMs displayed well-formed sarcomeres and their organization was similar between the two cell lines. However, cardiac 3D organoids generated with LMNA p.H222P hiPSC-CMs showed an impaired contractility compared to control organoids. Calcium transient recordings in LMNA p.H222P mutant cardiomyocytes showed a significantly higher calcium transient amplitude with a significantly slower calcium re-uptake. Transcriptomic analyses suggested a global mitochondrial dysfunction and in particular an impaired mitochondrial calcium uptake with a significantly decreased expression of the mitochondrial calcium uniporter (MCU). This decrease in MCU expression was confirmed by western blot and was accompanied by an increased MICU1:MCU, as well as an increased PDH Ser232 and PDH Ser300 phosphorylation, indicating a decreased mitochondrial calcium uptake in the LMNA mutant hiPSC-CMs. Measurement of mitochondrial respiration showed lower basal and maximal respiration in LMNA p.H222P hiPSC-CMs. Consistently, the ATP levels were significantly lower in LMNA p.H222P hiPSC-CMs as compared to isogenic controls.\u0000Conclusions: LMNA p.H222P mutant hiPSC-CMs exhibit contractile dysfunction associated with mitochondrial dysfunction with impaired MCU complex activity, decreased mitochondrial calcium homeostasis and reduced mitochondrial energy production.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"11 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142183097","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Deep Learning Framework for Predicting Prognostically Relevant Consensus Molecular Subtypes in HPV-Positive Cervical Squamous Cell Carcinoma from Routine Histology Images","authors":"Ruoyu Wang, Gozde Gunesli, Vilde Eide Skingen, Kari-Anne Frikstad Valen, Heidi Lyng, Lawrence Young, Nasir Rajpoot","doi":"10.1101/2024.08.16.608264","DOIUrl":"https://doi.org/10.1101/2024.08.16.608264","url":null,"abstract":"Despite efforts in human papillomavirus (HPV) prevention and screening, cervical cancer remains the fourth most prevalent cancer among women globally. In this study, we propose an end-to-end deep learning framework to investigate histological correlates of the two consensus molecular subtypes (CMS) of HPV-positive cervical squamous cell carcinoma (CSCC) patients. Analysing three international CSCC cohorts (n=545 patients), we demonstrate that the genomically determined CMS can be predicted from routine H&E-stained histology slides, with our Digital-CMS scores achieving significant patient stratifications in terms of disease-specific survival (TCGA p=0.0022, Oslo p=0.0495) and disease-free survival (TCGA p=0.0495, Oslo p=0.0282). In addition, our extensive analyses reveal distinct tumour microenvironment (TME) differences between the two CMS subtypes of the CSCC cohorts. Notably, CMS-C1 CSCC subgroup has markedly increased lymphocyte presence, whereas CMS-C2 subgroup has high nuclear pleomorphism, an elevated neutrophil-to-lymphocyte ratio, and increased neutrophil density. Analysis of representative histological regions reveals higher degree of malignancy in CMS-C2 patients, associated with poor prognosis. This study introduces a potentially clinically advantageous Digital-CMS score derived from digitised WSIs of routine H&E-stained tissue sections, offers new insights into TME differences impacting patient prognosis and potential therapeutic targets, and identifies histological patterns serving as potential surrogate markers of the two CMS subtypes for clinical application.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"8 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142183095","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid and Sensitive Detection of Fusarium oxysporum f. sp. cubense Tropical Race 4 Using a RPA-DETECTR Assay","authors":"Jelli Venkatesh, Joanna Jankowicz-Cieslak, Hassan Mduma, Mirta Matijevic, Adel Ali, Isabel Cristina Calle Balbin, Mauricio Soto-Suarez, Cinthya Zorrilla, Pooja Bhatnagar-Mathur","doi":"10.1101/2024.08.15.608054","DOIUrl":"https://doi.org/10.1101/2024.08.15.608054","url":null,"abstract":"Early and prompt detection of banana wilt pathogen Fusarium oxysporum f. sp. cubense (Foc), tropical race 4 (Foc TR4), causing global banana crop losses is crucial to curtail the disease spread, minimize damage and implement quarantine measures. We report the first successfully developed DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) assay for the highly sensitive and specific detection of Foc TR4, validated across several isolates. Notably, specific crRNA spacer sequences and recombinant polymerase amplification (RPA) primers designed and evaluated for the DETECTR assay exhibited exceptional sensitivity, detecting Foc TR4 with genomic DNA as low as 0.005 ng, and demonstrated a very high specificity, and reproducibility. The RPA-DETECTR assay was validated using a diverse panel of samples, including both target and non-target pathogens confirming its robustness and reliability across different types of samples, ensuring practical application under varied conditions. In terms of throughput, the RPA-DETECTR assays enabled faster detection of Foc TR4 than other molecular diagnostics including PCR, quantitative PCR (qPCR) and droplet digital PCR (ddPCR) analyses, making this assay suitable for point-of-care (POC) detection of Foc TR4, and facilitating rapid decision-making and immediate response. This rapid detection capability is crucial in restricting any potential transboundary movement as part of ongoing disease management efforts.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"173 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142182913","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}