Direct Detection of SARS CoV-2 by Practical Paper Based Imunoassay as an Instrument Free and Low Cost by CR3022

Yıl 2024, Cilt: 40 Sayı: 3, 528 - 534, 30.12.2024

Dilek Çam Derin

Öz

Pandemia could be emerged at any time and resulted with death in the world. Severe acute respiratory syndrome coronavirus 2 (SARS CoV-2) caused the latest pandemia. Because of the absence of rapid diagnostic assays pandemia has surged in and caused the death of lots of people as SARS CoV-2 appeared at first time. Although reverse transcriptase polymerase chain reaction (RT-PCR) accepted as the most reliable test was used for the viral detection at first time, there are shortcomings including the need of educated personnel and difficulty of the usage of this test in undeveloped countries. For this reason, developing the rapid assays for direct recognition of virus is always eesential. Paper based immunoassays are significant among these assays and they are known as dot-blot assay (DBA). It is practical and easy to use since they are suitable for direct application of body fluids. Antibodies and aptamers are the most important reagents of diagnostic assays and it should be used having the high affinity of them. In this work, DBA was prepared by antibody-aptamer based sandwich assay and reported the suitibility of it for viral detection.

Anahtar Kelimeler

Dot blot assay, viral diagnosis, paper assay, SARS CoV-2

CR3022 Antikoru ile Cihazdan Bağımsız Pratik Kağıt Tabanlı İmmünotest ile Direkt SARS CoV-2 Teşhisi

Öz

Pandemi dünyada her an ortaya çıkabilir ve birçok ölümle sonuçlanabilir. En son görülen pandemiye şiddetli akut solunum yolu sendromu koronavirüs 2 (SARS CoV-2) sebep olmuştur. SARS CoV-2 ilk kez ortaya çıktığı için hızlı tanı testlerinin yokluğu sebebiyle pandemi hızla ilerlemiş ve birçok kişinin ölümüne sebep olmuştur. Virüs tanısı için ilk olarak en güvenilir test olarak kabul edilen ters transkripsiyon polimeraz zincir reaksiyonu (RT-PZR) kullanılsa da eğitimli personel ihtiyacı duyması ve gelişmemiş ülkelerde bu testin uygulama zorluğu sebebiyle eksikleri vardır. Bu nedenle direkt virüs teşhisine yönelik hızlı testlerin geliştirilmesi her zaman elzemdir. Bu testler arasında kağıt tabanlı immünotestler önemlidir ve çoğunlukla dot-blot assay (noktasal test-NT) olarak bilinir. Vücut sıvılarının direkt uygulanmasına uygun olduğu için pratik ve kullanımı kolaydır. Antikor ve aptamerler tanı testlerinin en önemli bileşenleridir ve bunlardan afinitesi iyi olanlar kullanılmalıdır. Bu araştırmada antikor-aptamer sandviç testine dayalı olarak NT hazırlanmış ve viral teşhise uygun olup olmadığı bildirilmiştir.

Anahtar Kelimeler

Dot blot assay, viral diagnosis, paper assay, SARS CoV-2

Teşekkür

Çalışmada yardımları dokunan Dr. Irmak İÇEN TAŞKIN ve Eray TATLICI’ya teşekkür ederim

Kaynakça

• [1] Alafeef, M., Dighe, K., Moitra, P., Pan, D. 2020. Rapid, ultrasensitive, and quantitative detection of SARS- CoV-2 using antisense oligonucleotides directed electrochemical biosensor chip. ACS nano, 14:17028- 17045.
• [2] Ibrahim, N., Jamaluddin, N. D., Tan, L. L., Mohd Yusof, N. Y. 2021. A review on the development of gold and silver nanoparticles-based biosensor as a detection strategy of emerging and pathogenic RNA virus. Sensors, 21(15):5114.
• [3] Daniel, S. K., Pai, P. S., Sabbella, H. R., Singh, K., Rangaiah, A., Basawarajappa, S. G., Thakur, C. S. 2022. Handheld, low-cost, aptamer-based sensing device for rapid SARS-CoV-2 RNA detection using novelly synthesized gold nanoparticles. IEEE Sensors Journal, 22(19): 18437-18445.
• [4] Amanat, F., Stadlbauer, D., Strohmeier, S., Nguyen, T. H. O., Chromikova, V., McMahon, M., Jiang, K., Arunkumar, G. A., Jurczyszak, D., Polanco, J., Bermudez-Gonzalez, M., Kleiner, G., Aydillo, T., Miorin, L., Fierer, D. S., Lugo, L. A., Kojic, E. M., Stoever, J., Liu, S. T. H., Cunningham-Rundles, C., Krammer, F. 2020. A serological assay to detect SARS-CoV-2 seroconversion in humans. Nature medicine, 26(7):1033–1036.
• [5] Çam Derin, D., Gültekin, E., Gündüz, E., Otlu, B. 2024. Comparison of Six Aptamer-Aptamer Pairs on Rapid Detection of SARS-CoV-2 by Lateral Flow Assay. Journal of AOAC International, 107(3):464–470.211.
• [6] Antoine, D., Mohammadi, M., Vitt, M., Dickie, J. M., Jyoti, S. S., Tilbury, M. A., Johnson, P. A., Wawrousek, K. E., Wall, J. G. 2022. Rapid, Point-of-Care scFv-SERS Assay for Femtogram Level Detection of SARS-CoV-2. ACS sensors, 7(3):866–873.13.
• [7] Thippornchai, N., Pengpanich, S., Jaroenram, W., Kosoltanapiwat, N., Sukphopetch, P., Kiatpathomchai, W., Leaungwutiwong, P. 2024. A colorimetric reverse-transcription loop-mediated isothermal amplification method targeting the L452R mutation to detect the Delta variant of SARS-CoV-2. Scientific reports, 14(1):21961.
• [8] Lisi, S., Malerba, F., Quaranta, P., Florio, R., Vitaloni, O., Monaca, E., Bruni Ercole, B., Bitonti, A. R., Del Perugia, O., Mignanelli, M., Perrera, P., Sabbatella, R., Raimondi, F., Piazza, C. R., Moles, A., Alfano, C., Pistello, M., Cattaneo, A. 2024. Selection and characterization of human scFvs targeting the SARS-CoV-2 nucleocapsid protein isolated from antibody libraries of COVID-19 patients. Scientific reports, 14(1):15864.
• [9] Ge, S., Wu, R., Zhou, T., Liu, X., Zhu, J., Zhang, X. 2022. Specific anti-SARS-CoV-2 S1 IgY-scFv is a promising tool for recognition of the virus. AMB Express, 12(1):18.
• [10] Kim, H. Y., Lee, J. H., Kim, M. J., Park, S. C., Choi, M., Lee, W., Ku, K. B., Kim, B. T., Changkyun Park, E., Kim, H. G., Kim, S. I. 2021. Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins. Biosensors & bioelectronics, 175:112868.
• [11] Tohari, T. R., Anshori, I., Baroroh, U., Nugroho, A. E., Gumilar, G., Kusumawardani, S., Syahruni, S., Yuliarto, B., Arnafia, W., Faizal, I., Hartati, Y. W., Subroto, T., Yusuf, M. 2022. Development of a Single-Chain Variable Fragment of CR3022 for a Plasmonic-Based Biosensor Targeting the SARS-CoV-2 Spike Protein. Biosensors, 12(12):1133.
• [12] Kim, W., Song, E. S., Lee, S. H., Yang, S. H., Cho, J., Kim, S. J. 2024. A new DNA aptamer which binds to SARS-CoV-2 spike protein and reduces pro-inflammatory response. Scientific reports, 14(1):7516.
• [13] Yang, M., Li, C., Ye, G., Shen, C., Shi, H., Zhong, L., Tian, Y., Zhao, M., Wu, P., Hussain, A., Zhang, T., Yang, H., Yang, J., Weng, Y., Liu, X., Wang, Z., Gan, L., Zhang, Q., Liu, Y., Yang, G., Zhao, Y. 2024. Aptamers targeting SARS-CoV-2 nucleocapsid protein exhibit potential anti pan-coronavirus activity. Signal transduction and targeted therapy, 9(1):40.
• [14] Chen, Z., Wu, Q., Chen, J., Ni, X., Dai, J. 2020. A DNA Aptamer Based Method for Detection of SARS-CoV-2 Nucleocapsid Protein. Virologica Sinica, 35(3):351–354.20.
• [15] Yang, L. F., Kacherovsky, N., Panpradist, N., Wan, R., Liang, J., Zhang, B., Salipante, S. J., Lutz, B. R., Pun, S. H. 2022. Aptamer Sandwich Lateral Flow Assay (AptaFlow) for Antibody-Free SARS-CoV-2 Detection. Analytical chemistry, 94(20):7278–7285.
• [16] Erdem, A., Senturk, H., Yildiz, E., Maral, M. 2024. Optimized aptamer-based next generation biosensor for the ultra-sensitive determination of SARS-CoV-2 S1 protein in saliva samples. International journal of biological macromolecules, 281(Pt 1):136233.
• [17] Khan, R., Deshpande, A. S., Proteasa, G., Andreescu, S. 2024. Aptamer-based electrochemical biosensor with S protein binding affinity for COVID-19 detection: Integrating computational design with experimental validation of S protein binding affinity. Sensors and Actuators B: Chemical, 399:134775.
• [18] Kim, S., Lee, J. H. 2022. Current advances in paper-based biosensor technologies for rapid COVID-19 diagnosis. Biochip journal, 16(4):376-396.
• [19] Derin, D. C., Gültekin, E., Taşkın, I. İ., Yakupoğulları, Y. 2023. Development of nucleic acid based lateral flow assays for SARS-CoV-2 detection. Journal of bioscience and bioengineering, 135(2):87-92.
• [20] Najafabad, M. B., Rastin, S. J., Taghvaei, F., Khiyavi, A. A. 2022. A review on applications of gold nanoparticles-based biosensor for pathogen detection. Advances in Natural Sciences: Nanoscience and Nanotechnology, 13(3):033002.
• [21] Diamandis, E. P., Christopoulos, T. K. 1996. Immunoassay. Academic Press.
• [22] Apú, N., Madrigal-Redondo, G., Vega, M. H., Corrales-Aguilar, E., Segura-Ulate, I. 2023. Development of an instrument-free and low-cost ELISA dot-blot test to detect antibodies against SARS-CoV-2. Open life sciences, 18(1):20220577.
• [23] Norouzi, M., Truong, T., Jaenes, K., Warner, B. M., Vendramelli, R., Tierney, K., Kobasa, D., Tailor, N., Plant, P., Dos Santos, C., Babiuk, S., Ambagala, A., Pardee, K. 2023. Cell-Free Dot Blot: an Ultra-Low-Cost and Practical Immunoassay Platform for Detection of Anti-SARS-CoV-2 Antibodies in Human and Animal Sera. Microbiology spectrum, 11(2):e0245722. Advance online publication.215.
• [24] Celiker, T., Ghorbanizamani, F., Moulahoum, H., Guler Celik, E., Tok, K., Zihnioglu, F., Cicek, C., Sertoz, R., Arda, B., Goksel, T., Turhan, K., Timur, S., Yagci, Y. 2022. Fluorescent bioassay for SARS-CoV-2 detection using polypyrene-g-poly(ε-caprolactone) prepared by simultaneous photoinduced step-growth and ring- opening polymerizations. Mikrochimica acta, 189(5):202.
• [25] Connelly, G. G., Kirkland, O. O., Bohannon, S., Lim, D. C., Wilson, R. M., Richards, E. J., Tay, D. M., Jee, H., Hellinger, R. D., Hoang, N. K., Hao, L., Chhabra, A., Martin-Alonso, C., Tan, E. K. W., Koehler, A. N., Yaffe, M. B., London, W. B., Lee, P. Y., Krammer, F., Bohannon, R. C., Li, H. 2022. Direct capture of neutralized RBD enables rapid point-of-care assessment of SARS-CoV-2 neutralizing antibody titer. Cell reports methods, 2(8):100273.
• [26] Lee, J. H., Choi, M., Jung, Y., Lee, S. K., Lee, C. S., Kim, J., Kim, J., Kim, N. H., Kim, B. T., Kim, H. G. (2021). A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2). Biosensors & bioelectronics, 171:112715.
• [27] Singh, B., Datta, B., Ashish, A., Dutta, G. 2021. A comprehensive review on current COVID-19 detection methods: From lab care to point of care diagnosis. Sensors International, 2:100119.
• [28] Wang, J., Drelich, A. J., Hopkins, C. M., Mecozzi, S., Li, L., Kwon, G., Hong, S. 2022. Gold nanoparticles in virus detection: Recent advances and potential considerations for SARS‐CoV‐2 testing development. Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology, 14(1):e1754.
• [29] Tian, X., Li, C., Huang, A., Xia, S., Lu, S., Shi, Z., Lu, L., Jiang, S., Yang, Z., Wu, Y., Ying, T. 2020. Potent binding of 2019 novel coronavirus spike protein by a SARS coronavirus-specific human monoclonal antibody. Emerg. Microbes Infect, 9:382–385.
• [30] Song, Y., Song, J., Wei, X., Huang, M., Sun, M., Zhu, L., Lin, B., Shen, H., Zhu, Z., Yang, C. 2020. Discovery of Aptamers Targeting the Receptor-Binding Domain of the SARS-CoV-2 Spike Glycoprotein. Analytical chemistry, 92(14):9895–9900.
• [31] Huo, T. M., Peng, C. F., Xu, C. L., Liu, L. Q. 2006. Development of colloidalgold-based immunochromatographic assay for the rapid detection of medroxyprogesterone acetate residues. Food and Agricultural Immunology, 17:183-190.
• [32] Liu, R., Zhang, Y., Zhang, S., Qiu, W., Gao, Y. 2014. Silver enhancement of gold nanoparticles for biosensing: from qualitative to quantitative. Applied Spectroscopy Reviews, 49(2):121-138.
• [33] Li, J., Zhang, Z., Gu, J., Stacey, H. D., Ang, J. C., Capretta, A., Filipe, C. D. M., Mossman, K. L., Balion, C., Salena, B. J., Yamamura, D., Soleymani, L., Miller, M. S., Brennan, J. D., Li, Y. 2021. Diverse high-affinity DNA aptamers for wild-type and B.1.1.7 SARS-CoV-2 spike proteins from a pre-structured DNA library. Nucleic acids research, 49(13):7267–7279.