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Tests basés sur CRISPR pour la détection des infections par les virus BK et JC après transplantation rénale

  • role： Contribute equally, First author同等贡献者, 第一作者
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Yu Liu1,  
  • role： Contribute equally同等贡献者
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Jing-Song Xu1,  
  • role： Contribute equally同等贡献者
  • Affiliation：

    Department of Pathology, the Fifth People’s Hospital of Shanghai, Fudan University, Shanghai 200240, China

Li Cao2,  
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Shuang Yang1,  
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Tian-Ming Li1,  
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Hai-Qian Huang1,  
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Jun-Heng Zhang1,  
  • Affiliation：

    Central Laboratory, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Xue Zhao3,  
  • role： Corresponding author通信作者
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

  • Email：qq2005011@163.com
  • Introduction：* Qian Liu qq2005011@163.com
Qian Liu1*,  
  • role： Corresponding author通信作者
  • Affiliation：

    Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China

  • Email：lishun86@126.com
  • Introduction：Shun Li lishun86@126.com
Shun Li4*,  
  • role： Corresponding author通信作者
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

  • Email：rjlimin@shsmu.edu.cn
  • Introduction：Min Li rjlimin@shsmu.edu.cn
Min Li1*,  
  • role： Corresponding author通信作者
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

  • Email：renjijywh@shsmu.edu.cn
  • Introduction：Hua Wang renjijywh@shsmu.edu.cn
Hua Wang1*

résumé

Les receveurs de transplantation d’organes sont confrontés à des risques importants d’infections aiguës ou chroniques qui menacent la survie du greffon. Le virus BK (BKV) et le virus JC (JCV) sont deux virus opportunistes importants, susceptibles de provoquer une néphropathie associée au polyomavirus et une perte du rein greffé chez les patients en état d’immunosuppression après une transplantation rénale. Par conséquent, la détection en temps opportun et la surveillance continue de la charge virale sont indispensables. Cependant, les méthodes diagnostiques actuelles restent limitées, et le développement de nouvelles technologies de détection moléculaire est extrêmement urgent. Les séquences et concentrations de l’ARN CRISPR (crRNA), la concentration de Cas13a et les amorces pour l’amplification par recombinase polymérase (RPA) ont été optimisées pour la détection de BKV et JCV. Ensuite, une nouvelle puce microfluidique à double goutte a été conçue et fabriquée, puis intégrée avec CRISPR (ddCRISPR) pour détecter simultanément qualitativement BKV et JCV. Par la suite, le test ddCRISPR a été validé avec des échantillons cliniques. Ensuite, une bande de flux latéral combinée au CRISPR (LFCRISPR) a été développée pour la détection de BKV et JCV dans des environnements à ressources limitées. Un système de réaction RPA-CRISPR en un seul pot a été établi et optimisé pour la détection de BKV et JCV. Le ddCRISPR peut détecter simultanément et rapidement BKV et JCV avec une grande sensibilité (10 copies/ml pour BKV et 1 copie/ml pour JCV), et fournir une quantification absolue, ce qui convient à la détection de la charge virale et favorise un traitement personnalisé et précis des receveurs de transplantation d’organes. LFCRISPR simplifie le processus opérationnel grâce à une lecture visuelle simple, facilitant le dépistage viral après transplantation d’organe. Ces plateformes intègrent le test moléculaire dans le modèle de traitement de transplantation, réduisant ainsi les coûts, prolongeant la durée de survie du greffon, améliorant les résultats cliniques de la prise en charge postopératoire lors de la transplantation rénale, et améliorant la qualité de vie des patients.

mots-clés

virus BK (BKV); virus JC (JCV); transplantation rénale; répétitions palindromiques courtes régulièrement espacées (CRISPR); microfluidique; test à flux latéral (LFA)

References

  1. 1..
    Pinto M,Dobson S. BK and JC virus: a review. J Infect. 2014;68(Suppl 1):S2-8.
  2. 2..
    Mazzoni E,Pellegrinelli E,Mazziotta C,Lanzillotti C,Rotondo JC,Bononi I, et al. Mother-to-child transmission of oncogenic polyomaviruses BKPyV, JCPyV and SV40. J Infect. 2020;80(5):563-70.
  3. 3..
    Bauman Y,Nachmani D,Vitenshtein A,Tsukerman P,Drayman N,Stern-Ginossar N, et al. An identical miRNA of the human JC and BK polyoma viruses targets the stress-induced ligand ULBP3 to escape immune elimination. Cell Host Microbe. 2011;9(2):93-102.
  4. 4..
    Hirsch HH,Steiger J. Polyomavirus BK. Lancet Infect Dis.2003;3(10):611-23.
  5. 5..
    Olson A,Lin R,Marin D,Rafei H,Bdaiwi MH,Thall PF, et al. Third-party BK virus-specific cytotoxic t lymphocyte therapy for hemorrhagic cystitis following allotransplantation. J Clin Oncol. 2021;39(24):2710-9.
  6. 6..
    Cortese I,Reich DS,Nath A. Progressive multifocal leukoencepha-lopathy and the spectrum of JC virus-related disease. Nat Rev Neurol.2021;17(1):37-51.
  7. 7..
    Cortese I,Muranski P,Enose-Akahata Y,Ha SK,Smith B,Monaco M, et al. Pembrolizumab treatment for progressive multifocal leukoencephalopa-thy. N Engl J Med. 2019;380(17):1597-605.
  8. 8..
    Major EO,Yousry TA,Clifford DB. Pathogenesis of progressive multifocal leukoencephalopathy and risks associated with treatments for multiple sclerosis: a decade of lessons learned. Lancet Neurol. 2018;17(5):467-80.
  9. 9..
    Seppälä HM,Helanterä IT,Laine PKS,Lautenschlager IT,Paulín LG,Jah-nukainen TJ, et al. Archetype JC polyomavirus (JCPyV) prevails in a rare case of JCPyV nephropathy and in stable renal transplant recipients with JCPyV viruria. J Infect Dis. 2017;216(8):981-9.
  10. 10..
    Peretti A,Geoghegan EM,Pastrana DV,Smola S,Feld P,Sauter M, et al.Characterization of BK polyomaviruses from kidney transplant recipients suggests a role for APOBEC3 in driving in-host virus evolution. Cell Host Microbe. 2018;23(5):628-35.e7.
  11. 11..
    Hariharan S. BK virus nephritis after renal transplantation. Kidney Int.2006;69(4):655-62.
  12. 12..
    Brennan DC,Agha I,Bohl DL,Schnitzler MA,Hardinger KL,Lockwood M, et al. Incidence of BK with tacrolimus versus cyclosporine and impact of preemptive immunosuppression reduction. Am J Transplant.2005;5(3):582-94.
  13. 13..
    Tan CS,Koralnik IJ. Progressive multifocal leukoencephalopathy and other disorders caused by JC virus: clinical features and pathogenesis. Lancet Neurol. 2010;9(4):425-37.
  14. 14..
    Hirsch HH,Knowles W,Dickenmann M,Passweg J,Klimkait T,Mihatsch MJ, et al. Prospective study of polyomavirus type BK replication and nephropathy in renal-transplant recipients. N Engl J Med.2002;347(7):488-96.
  15. 15..
    Nankivell BJ,Renthawa J,Shingde M,Khan A. The importance of kidney medullary tissue for the accurate diagnosis of BK virus allograft nephrop-athy. Clin J Am Soc Nephrol. 2020;15(7):1015-23.
  16. 16..
    Li C,Hughes C,Ding R,Snopkowski C,Salinas T,Schwartz J, et al. Develop-ment of a Bak gene based standard curve for absolute quantification of BK virus in real time quantitative PCR assay and noninvasive diagnosis of BK virus nephropathy in kidney allograft recipients. J Immunol Methods.2022;509:113341.
  17. 17..
    Bateman AC,Greninger AL,Atienza EE,Limaye AP,Jerome KR,Cook L.Quantification of BK virus standards by quantitative real-time PCR and droplet digital PCR is confounded by multiple virus populations in the WHO BKV international standard. Clin Chem. 2017;63(3):761-9.
  18. 18..
    Kevadiya BD,Machhi J,Herskovitz J,Oleynikov MD,Blomberg WR,Bajwa N, et al. Diagnostics for SARS-CoV-2 infections. Nat Mater.2021;20(5):593-605.
  19. 19..
    Lobato IM,O’sullivan CK. Recombinase polymerase amplification: basics, applications and recent advances. Trends Analyt Chem. 2018;98:19-35.
  20. 20..
    Hun X,Meng Y. Electron acceptors co-regulated self-powered photo-electrochemical strategy and its application for circulating tumor nucleic acid detection coupled with recombinase polymerase amplification. Anal Chem. 2020;92(17):11771-8.
  21. 21..
    Yin W,Zhuang J,Li J,Xia L,Hu K,Yin J, et al. Digital recombinase polymer-ase amplification, digital loop-mediated isothermal amplification, and digital CRISPR-Cas assisted assay: current status, challenges, and perspec-tives. Small. 2023;19(49):e2303398.
  22. 22..
    Tomar S,Lavickova B,Guiducci C. Recombinase polymerase amplification in minimally buffered conditions. Biosens Bioelectron. 2022;198:113802.
  23. 23..
    Patchsung M,Jantarug K,Pattama A,Aphicho K,Suraritdechachai S,Mee-sawat P, et al. Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA. Nat Biomed Eng. 2020;4(12):1140-9.
  24. 24..
    Gootenberg JS,Abudayyeh OO,Lee JW,Essletzbichler P,Dy AJ,Joung J, et al. Nucleic acid detection with CRISPR-Cas13a/C2c2. Science.2017;356(6336):438-42.
  25. 25..
    Chen JS,Ma E,Harrington LB,Da Costa M,Tian X,Palefsky JM, et al.CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity. Science. 2018;360(6387):436-9.
  26. 26..
    Gootenberg JS,Abudayyeh OO,Kellner MJ,Joung J,Collins JJ,Zhang F.Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018;360(6387):439-44.
  27. 27..
    Fan XW,Gao ZF,Ling DD,Wang DH,Cui Y,Du HQ, et al. CRISPR/Cas9 nickase mediated signal amplification integrating with the trans-cleavage activity of Cas12a for highly selective and sensitive detection of single base mutations. Mil Med Res. 2024;11(1):25.
  28. 28..
    Li ZH,Wang J,Xu JP,Wang J,Yang X. Recent advances in CRISPR-based genome editing technology and its applications in cardiovascular research. Mil Med Res. 2023;10(1):12.
  29. 29..
    Zhou S,Dong J,Deng L,Wang G,Yang M,Wang Y, et al. Endonuclease-assisted PAM-free recombinase polymerase amplification coupling with CRISPR/Cas12a (E-PfRPA/Cas) for sensitive detection of DNA methylation.ACS Sens. 2022;7(10):3032-40.
  30. 30..
    Kaminski MM,Alcantar MA,Lape IT,Greensmith R,Huske AC,Valeri JA, et al. A CRISPR-based assay for the detection of opportunistic infections post-transplantation and for the monitoring of transplant rejection. Nat Biomed Eng. 2020;4(6):601-9.
  31. 31..
    Liu Y,Xu J,Chen X,Yang S,Li J,Zhou H, et al. Digital CRISPR/Cas13a-RPA based assay for rapid and sensitive detection of JC virus in renal trans-plant patients. Microchem J. 2024;196:109655.
  32. 32..
    Decaprio JA,Garcea RL. A cornucopia of human polyomaviruses. Nat Rev Microbiol. 2013;11(4):264-76.
  33. 33..
    Tzannou I,Papadopoulou A,Naik S,Leung K,Martinez CA,Ramos CA, et al. Off-the-shelf virus-specific T cells to treat BK virus, human herpes-virus 6, cytomegalovirus, Epstein-Barr virus, and adenovirus infections after allogeneic hematopoietic stem-cell transplantation. J Clin Oncol.2017;35(31):3547-57.
  34. 34..
    Wekerle T,Segev D,Lechler R,Oberbauer R. Strategies for long-term preservation of kidney graft function. Lancet. 2017;389(10084):2152-62.
  35. 35..
    Sumida K,Nadkarni GN,Grams ME,Sang Y,Ballew SH,Coresh J, et al.Conversion of urine protein-creatinine ratio or urine dipstick protein to urine albumin-creatinine ratio for use in chronic kidney disease screening and prognosis: an individual participant-based meta-analysis. Ann Intern Med. 2020;173(6):426-35.
  36. 36..
    Abdou Mohamed MA,Kozlowski HN,Kim J,Zagorovsky K,Kantor M,Feld JJ, et al. Diagnosing antibiotic resistance using nucleic acid enzymes and gold nanoparticles. ACS Nano. 2021;15(6):9379-90.
  37. 37..
    Daunay A,Duval A,Baudrin LG,Buhard O,Renault V,Deleuze JF, et al.Low temperature isothermal amplification of microsatellites drastically reduces stutter artifact formation and improves microsatellite instability detection in cancer. Nucleic Acids Res. 2019;47(21):e141.
  38. 38..
    Mohsen MG,Kool ET. The discovery of rolling circle amplification and rolling circle transcription. Acc Chem Res. 2016;49(11):2540-50.
  39. 39..
    Soares RRG,Madaboosi N,Nilsson M. Rolling circle amplification in inte-grated microsystems: an uncut gem toward massively multiplexed path-ogen diagnostics and genotyping. Acc Chem Res. 2021;54(21):3979-90.
  40. 40..
    Ludwig KU,Schmithausen RM,Li D,Jacobs ML,Hollstein R,Blumenstock K, et al. LAMP-seq enables sensitive, multiplexed COVID-19 diagnostics using molecular barcoding. Nat Biotechnol. 2021;39(12):1556-62.
  41. 41..
    Song M,Hong S,Lee LP. Multiplexed ultrasensitive sample-to-answer RT-LAMP chip for the identification of SARS-CoV-2 and influenza viruses. Adv Mater. 2023;35(10):e2207138.
  42. 42..
    Bi S,Yue S,Zhang S. Hybridization chain reaction: a versatile molecu-lar tool for biosensing, bioimaging, and biomedicine. Chem Soc Rev.2017;46(14):4281-98.
  43. 43..
    Arshad F,Abdillah AN,Shivanand P,Ahmed MU. CeO2 nanozyme medi-ated RPA/CRISPR-Cas12a dual-mode biosensor for detection of invA gene in Salmonella. Biosens Bioelectron. 2024;247:115940.
  44. 44..
    Lin C,Chen F,Huang D,Li W,He C,Tang Y, et al. A universal all-in-one RPA-Cas12a strategy with de novo autodesigner and its application in on-site ultrasensitive detection of DNA and RNA viruses. Biosens Bioelectron.2023;239:115609.
  45. 45..
    Ahamed MA,Khalid MaU,Dong M,Politza AJ,Zhang Z,Kshirsagar A, et al. Sensitive and specific CRISPR-Cas12a assisted nanopore with RPA for Monkeypox detection. Biosens Bioelectron. 2024;246:115866.
  46. 46..
    Deng R,Zhang K,Li J. Isothermal amplification for microRNA detection:from the test tube to the cell. Acc Chem Res. 2017;50(4):1059-68.
  47. 47..
    Nyaruaba R,Mwaliko C,Dobnik D,Neužil P,Amoth P,Mwau M, et al.Digital PCR applications in the SARS-CoV-2/COVID-19 era: a roadmap for future outbreaks. Clin Microbiol Rev. 2022;35(3):e0016821.
  48. 48..
    Albayrak C,Jordi CA,Zechner C,Lin J,Bichsel CA,Khammash M, et al.Digital quantification of proteins and mRNA in single mammalian cells.Mol Cell. 2016;61(6):914-24.
  49. 49..
    Tian T,Shu B,Jiang Y,Ye M,Liu L,Guo Z, et al. An ultralocalized Cas13a assay enables universal and nucleic acid amplification-free single-mole-cule RNA diagnostics. ACS Nano. 2021;15(1):1167-78.
  50. 50..
    Zhao Y,Chen D,Xu Z,Li T,Zhu J,Hu R, et al. Integrating CRISPR-Cas12a into a microfluidic dual-droplet device enables simultaneous detection of HPV16 and HPV18. Anal Chem. 2023;95(6):3476-85.
  51. 51..
    Jin Z,Dong Z,Zhao X,Hang X,Lu Y,Zhang Q, et al. Sensitive, rapid detection of NCOA4-m6A towards precisely quantifying radiation dosage on a Cas13a-microdroplet platform. Biosens Bioelectron. 2023;242:115753.
  52. 52..
    Su G,Zhu M,Li D,Xu M,Zhu Y,Zhang Y, et al. Multiplexed lateral flow assay integrated with orthogonal CRISPR-Cas system for SARS-CoV-2 detection. Sens Actuators B Chem. 2022;371:132537.

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