اخبار و اعلانات

 آمار

تعداد نشریات286
تعداد نشریات سازمان84
تعداد شماره‌ها3,021
تعداد مقالات33,767
تعداد مشاهده مقاله45,090,526
تعداد دریافت فایل اصل مقاله50,945,279
Golchinfar, Fariba, Madani, Rasool, Hezarosi, Mahsa, Emami, Tara, Ghanizade, Arash. (1403). Development of an ELISA for SARS-CoV-2 Detection focusing on antibodies against nucleocapsid protein. سامانه مدیریت نشریات علمی, 80(1), 85-92. doi: 10.32592/ARI.2025.80.1.85
Fariba Golchinfar; Rasool Madani; Mahsa Hezarosi; Tara Emami; Arash Ghanizade. "Development of an ELISA for SARS-CoV-2 Detection focusing on antibodies against nucleocapsid protein". سامانه مدیریت نشریات علمی, 80, 1, 1403, 85-92. doi: 10.32592/ARI.2025.80.1.85
Golchinfar, Fariba, Madani, Rasool, Hezarosi, Mahsa, Emami, Tara, Ghanizade, Arash. (1403). 'Development of an ELISA for SARS-CoV-2 Detection focusing on antibodies against nucleocapsid protein', سامانه مدیریت نشریات علمی, 80(1), pp. 85-92. doi: 10.32592/ARI.2025.80.1.85
Golchinfar, Fariba, Madani, Rasool, Hezarosi, Mahsa, Emami, Tara, Ghanizade, Arash. Development of an ELISA for SARS-CoV-2 Detection focusing on antibodies against nucleocapsid protein. سامانه مدیریت نشریات علمی, 1403; 80(1): 85-92. doi: 10.32592/ARI.2025.80.1.85

Development of an ELISA for SARS-CoV-2 Detection focusing on antibodies against nucleocapsid protein

Archives of Razi Institute
مقاله 10، دوره 80، شماره 1، فروردین و اردیبهشت 2025، صفحه 85-92    اصل مقاله (358.73 K)
نوع مقاله: Original Articles
شناسه دیجیتال (DOI): 10.32592/ARI.2025.80.1.85
نویسندگان
Fariba Golchinfar1؛ Rasool Madani* 1؛ Mahsa Hezarosi1؛ Tara Emami2؛ Arash Ghanizade1
1Department of Proteomics and Biochemistry, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran
2Razi Vaccine and Serum Research Institute
چکیده
In late 2019, a new viral disease called COVID-19 caused by SARS-CoV-2 started in China and soon became a pandemic. This virus hurts human health and brings about financial damage to human societies, so researchers are trying to better identify and control it faster. Using the ELISA method to screen a large number of patients is always very helpful in infectious epidemics.
In this research, the nucleocapsid protein (NP) of the SARS-CoV-2 virus was used to measure serum antibodies, which was received from Avicenna Research Institute. We coated this antigen on each well of the plates of the ELISA test. After that, we added serum samples from medical diagnostic laboratories (positive and negative serums measured by ELISA and PCR. For this purpose, checkerboard titration for all serum samples and antigens was carried out to optimize ELISA. Our ELISA test was an indirect assay that could detect antibodies against nucleocapsid protein. Finally, we measured the cut-off, sensitivity, and specificity of our ELISA test.
Our results showed that 95% sensitivity and 92% specificity. Also, in the intra-assay, the value of CV was 0.263%and in the inter-assay, the value of CV was 0.41%. As a result, it can be stated that this ELISA test was highly accurate and had high durability.
In conclusion, our kit showcases promising capabilities in the accurate and efficient detection of antibodies targeted against NP. This innovative approach not only enhances diagnostic accuracy but also holds significant potential for advancing antibody detection methodologies in the field of virology and immunology.
کلیدواژه‌ها
SARS-CoV-2؛ nucleocapsid protein؛ ELISA؛ COVID-19؛ diagnosis
عنوان مقاله [English]
طراحی کیت الایزای جدید علیه NP کرونا ویروس
چکیده [English]
در اواخر سال 2019، یک بیماری ویروسی جدید به نام COVID-19 ناشی از SARS-CoV-2 در چین شروع شد و به یک بیماری همه گیر تبدیل شد. این ویروس به سلامت انسان آسیب می رساند و آسیب مالی به جوامع بشری وارد می کند، بنابراین محققان در تلاش برای شناسایی و کنترل سریعتر آن هستند. استفاده از روش الایزا برای غربالگری تعداد زیادی از بیماران همیشه در اپیدمی های عفونی بسیار مفید واقع شده است.
در این تحقیق از پروتئین نوکلئوکپسید (NP) ویروس SARS-CoV-2 برای اندازه گیری آنتی بادی های سرم استفاده شد که از موسسه تحقیقات ابن سینا دریافت شد. ما این آنتی ژن را روی هر چاهک پلیت الایزا پوشاندیم. پس از آن، نمونه‌های سرم آزمایشگاه‌های تشخیص پزشکی (سرم‌های مثبت و منفی اندازه‌گیری شده با روش ELISA و PCR) را اضافه کردیم. بدین منظور، تیتراسیون تمام نمونه‌ها و آنتی‌ژن‌های سرم برای بهینه‌سازی الایزا انجام شد. در نهایت cut-off، حساسیت و ویژگی تست الایزا را اندازه گیری کردیم.
نتایج ما نشان داد که حساسیت 95 درصد و ویژگی 92 درصد است. همچنین تکرارپذیری این تست در یک ران کاری، به مقدار CV 0.263% و در ران های مختلف کاری، به مقدار CV 0.41% بود. در نتیجه می توان بیان کرد که این تست الایزا از دقت بالایی برخوردار بوده و ماندگاری بالایی داشته است.
در نتیجه، کیت ما قابلیت‌های امیدوارکننده‌ای را در تشخیص دقیق و کارآمد آنتی‌بادی‌های علیه NP نشان می‌دهد. این رویکرد نوآورانه نه تنها دقت تشخیصی را افزایش می‌دهد، بلکه پتانسیل قابل توجهی برای پیشبرد روش‌های تشخیص آنتی‌بادی در زمینه ویروس‌شناسی و ایمونولوژی دارد.
کلیدواژه‌ها [English]
کووید 19, پروتئین نوکلئوکپسید, الایزا, تشخیص
مراجع
  1. Yuki K, Fujiogi M, Koutsogiannaki S. COVID-19 pathophysiology: A review. Clinical Immunology. 2020;215:108427.
  2. Ciotti M, Ciccozzi M, Terrinoni A, Jiang W-C, Wang C-B, Bernardini S. The COVID-19 pandemic. Critical reviews in clinical laboratory sciences. 2020;57(6):365-88.
  3. Zhu N, Zhang D, Wang W, Li X, Yang B, Song J, et al. A novel coronavirus from patients with pneumonia in China, 2019. New England journal of medicine. 2020;382(8):727-33.
  4. The species Severe acute respiratory syndrome-related coronavirus: classifying 2019-nCoV and naming it SARS-CoV-2. Nature microbiology. 2020;5(4):536-44.
  5. Sidiq Z, Hanif M, Dwivedi KK, Chopra KK. Benefits and limitations of serological assays in COVID-19 infection. Indian Journal of Tuberculosis. 2020;67(4, Supplement):S163-S6.
  6. Høeg TB, Prasad V. Rapid antigen testing for COVID-19: Decreasing diagnostic reliability, potential detrimental effects and a lack of evidence to support continued public funding of community-based testing. Public Health Pract (Oxf). 2023;6:100451.
  7. Che X-y, Qiu L-w, Liao Z-y, Wang Y-d, Wen K, Pan Y-x, et al. Antigenic cross-reactivity between severe acute respiratory syndrome—associated coronavirus and human coronaviruses 229E and OC43. Journal of Infectious Diseases. 2005;191(12):2033-7.
  8. Dijkman R, Jebbink MF, Gaunt E, Rossen JW, Templeton KE, Kuijpers TW, et al. The dominance of human coronavirus OC43 and NL63 infections in infants. Journal of Clinical Virology. 2012;53(2):135-9.
  9. Wang Y, Sun S, Shen H, Jiang L, Zhang M, Xiao D, et al. Cross-reaction of SARS-CoV antigen with autoantibodies in autoimmune diseases. Cell Mol Immunol. 2004;1(4):304-7.
  10. Carattoli A, Bonito PD, Grasso F, Giorgi C, Blasi F, Niedrig M, et al. Recombinant protein‐based ELISA and immuno‐cytochemical assay for the diagnosis of SARS. Journal of medical virology. 2005;76(2):137-42.
  11. Guan M, Chen HY, Foo SY, Tan Y-J, Goh P-Y, Wee SH. Recombinant protein-based enzyme-linked immunosorbent assay and immunochromatographic tests for detection of immunoglobulin G antibodies to severe acute respiratory syndrome (SARS) coronavirus in SARS patients. Clinical and Vaccine Immunology. 2004;11(2):287-91.
  12. Leung DTM, Chi Hang TF, Chun Hung M, Sheung Chan PK, Cheung JLK, Niu H, et al. Antibody response of patients with severe acute respiratory syndrome (SARS) targets the viral nucleocapsid. The Journal of infectious diseases. 2004;190(2):379-86.
  13. Liu X, Shi Y, Li P, Li L, Yi Y, Ma Q, et al. Profile of antibodies to the nucleocapsid protein of the severe acute respiratory syndrome (SARS)-associated coronavirus in probable SARS patients. Clinical and vaccine immunology. 2004;11(1):227-8.
  14. Shi Y, Yi Y, Li P, Kuang T, Li L, Dong M, et al. Diagnosis of severe acute respiratory syndrome (SARS) by detection of SARS coronavirus nucleocapsid antibodies in an antigen-capturing enzyme-linked immunosorbent assay. Journal of clinical microbiology. 2003;41(12):5781-2.
  15. Woo PC, Lau SK, Tsoi H-w, Chan K-h, Wong BH, Che X-y, et al. Relative rates of non-pneumonic SARS coronavirus infection and SARS coronavirus pneumonia. The Lancet. 2004;363(9412):841-5.
  16. He Y, Zhou Y, Wu H, Kou Z, Liu S, Jiang S. Mapping of antigenic sites on the nucleocapsid protein of the severe acute respiratory syndrome coronavirus. J Clin Microbiol. 2004;42(11):5309-14.
  17. Masters PS, Sturman LS. Background paper. Functions of the coronavirus nucleocapsid protein. Adv Exp Med Biol. 1990;276:235-8.
  18. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193(1):265-75.
  19. Golchinfar F, Madani R, Hezarosi M. Study the Profile of Foot-and-mouth Disease Virus Protein by Electrophoresis and Identification of the Immunodominant Proteins. Archives of Razi Institute. 2023;78(5):1563-71.
  20. Madani R, Hezarosi M, Golchinfar F. Unveiling indirect ELISA test against nucleoprotein of H9N2 comparing with hemagglutination inhibition test. Archives of Razi Institute. 2024:-.
  21. Yüce M, Filiztekin E, Özkaya KG. COVID-19 diagnosis —A review of current methods. Biosensors and Bioelectronics. 2021;172:112752.
  22. Rai P, Kumar BK, Deekshit VK, Karunasagar I, Karunasagar I. Detection technologies and recent developments in the diagnosis of COVID-19 infection. Applied Microbiology and Biotechnology. 2021;105(2):441-55.
  23. Zhai P, Ding Y, Wu X, Long J, Zhong Y, Li Y. The epidemiology, diagnosis and treatment of COVID-19. International Journal of Antimicrobial Agents. 2020;55(5):105955.
  24. Zhao L, Han N, Zheng Y, Rao H, Li J, Chen Y, et al. Detection of Antibodies Against the SARS-CoV-2 Spike Protein and Analysis of the Peripheral Blood Mononuclear Cell Transcriptomic Profile, 15 Years After Recovery From SARS. 2021;11.
  25. Tan YJ, Goh PY, Fielding BC, Shen S, Chou CF, Fu JL, et al. Profiles of antibody responses against severe acute respiratory syndrome coronavirus recombinant proteins and their potential use as diagnostic markers. Clinical and diagnostic laboratory immunology. 2004;11(2):362-71.
  26. Bastos ML, Tavaziva G, Abidi SK, Campbell JR, Haraoui L-P, Johnston JC, et al. Diagnostic accuracy of serological tests for covid-19: systematic review and meta-analysis. bmj. 2020;370.
  27. Böger B, Fachi MM, Vilhena RO, Cobre AF, Tonin FS, Pontarolo R. Systematic review with meta-analysis of the accuracy of diagnostic tests for COVID-19. American journal of infection control. 2021;49(1):21-9.
  28. Caini S, Bellerba F, Corso F, Díaz-Basabe A, Natoli G, Paget J, et al. Meta-analysis of diagnostic performance of serological tests for SARS-CoV-2 antibodies up to 25 April 2020 and public health implications. Eurosurveillance. 2020;25(23):2000980.
  29. Deeks J, Dinnes J, Takwoingi Y, Davenport C, Spijker R, Taylor-Phillips S, et al. Ruffano LFd, Harris IM, Price MJ, Dittrich S, Emperador D, Hooft L, Leeflang MM, Bruel AVd, Cochrane COVID-19 Diagnostic Test Accuracy Group. Antibody tests for identification of current and past infection with SARS-CoV-2. Cochrane Database Syst Rev. 2020.
  30. Ng KW, Faulkner N, Cornish GH, Rosa A, Harvey R, Hussain S, et al. Preexisting and de novo humoral immunity to SARS-CoV-2 in humans. Science. 2020;370(6522):1339-43.
  31. Tso FY, Lidenge SJ, Peña PB, Clegg AA, Ngowi JR, Mwaiselage J, et al. High prevalence of pre-existing serological cross-reactivity against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in sub-Saharan Africa. International Journal of Infectious Diseases. 2021;102:577-83.
  32. Tozetto-Mendoza TR, Kanunfre KA, Vilas-Boas LS, Sanchez Espinoza EP, Paião HGO, Rocha MC, et al. Nucleoprotein-based ELISA for detection of SARS-COV-2 IgG antibodies: Could an old assay be suitable for serodiagnosis of the new coronavirus? Journal of Virological Methods. 2021;290:114064.
  33. Chang C-k, Sue S-C, Yu T-h, Hsieh C-M, Tsai C-K, Chiang Y-C, et al. Modular organization of SARS coronavirus nucleocapsid protein. 2006;13:59-72.
آمار
تعداد مشاهده مقاله: 193
تعداد دریافت فایل اصل مقاله: 309


counter
سامانه مدیریت نشریات علمی. طراحی و پیاده سازی از سیناوب