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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
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