Günümüz Pandemisi COVID-19’un Laboratuvar Tanı Yöntemleri

Yıl 2020, Cilt: 3 Sayı: COVID-19, 111 - 115, 13.07.2020

Dilek Dülger , Seda Ekici

Öz

Bu derleme, Çin’in Hubei eyaletinin Wuhan şehrinde yeni bir korona virüs türünden (SARS-CoV-2) kaynaklanan vekısa bir süre içinde pandemiye sebep olan, Şiddetli Akut Solunum Sendromu Koronavirüs Hastalığı 19 (Covid-19)’ unlaboratuvar teşhis metotları ile ilgili bilgi vermek amacıyla hazırlanmıştır. Global olarak ciddi bir tahribata yol açanCovid-19 pandemisini önleyebilmek adına, hassas ve spesifik laboratuvar teşhis metotlarının geliştirilmesi, vakalarınhızlı ve güvenilir şekilde tanımlanabilmesi için oldukça önemlidir. Tüm viral hastalıkların tanısında olguğu gibi Covid-19’un tanısı için de moleküler ve serolojik testler kullanılmaktadır. Doğru ve güvenilir test sonuçları için, serolojikyanıt ve hastalığın evresi göz önünde bulundurularak, hangi testin yapılması gerektiğine karar verilmektedir. Virüs pikkonsantrasyonuna enfeksiyon başladıktan sonraki 5. günde ulaşmaktadır. Bu nedenle virüs, hastalığın başlangıcındansonraki yedi güne kadar nazofarenksten alınan örneklerde, PCR testi ile kolayca tespit edilebilir. Ülkemizde RNA’yabağlı RNA polimeraz (RdRp) gen fragmanını hedefleyen gerçek zamanlı PCR tabanlı analiz metodu, Covid-19 hastalığınıteşhis etmek üzere rutin olarak kullanılmaktadır. Covid-19 tanısına yönelik PCR tabanlı analiz işlemleri için idealnumuneler; nazofarenks ve/veya orofarenksten alınan sürüntü örnekleri ve pnömonili hastalarda bronkoalveolarlavaj sıvısı ile balgam örnekleridir. Ayrıca, Covid-19’un serolojik olarak teşhis edilebilmesi için spike proteinlerinin verekombinant nükleokapsid proteini tespitini sağlayan ELISA kitleri ve nazofarengeal sekresyonlardaki antikorların yada viral bileşenlerin tespiti amacıyla geliştirilen hızlı tanı kitleri kullanılmaktadır. Dünya Sağlık Örgütü (DSÖ) Covid-19tanısında hızlı antijen ve antikor testlerinin kullanımını sürveyans çalışmaları için tavsiye etmektedir.

Anahtar Kelimeler

COVID-19, Moleküler, SARS-Cov-2, Serolojik

Laboratory Diagnosis Methods of Today’s Pandemic COVID-19

Öz

This review is based on the laboratory diagnostic methods of Severe Acute Respiratory Syndrome CoronavirusDisease 19 (Covid-19) caused by a new type of corona virus (SARS-CoV-2) in Wuhan, Hubei province, China, causing apandemic in a short time. It is prepared to provide relevant information. In order to prevent the Covid-19 pandemic,which causes a serious global destruction, the development of sensitive and specific laboratory diagnostic methodsis crucial to identify cases quickly and reliably. Molecular and serological tests are used for the diagnosis of Covid-19as in the diagnosis of all viral diseases. For accurate and reliable test results, it is decided which test should be doneby considering the serological response and the stage of the disease. The virus reaches its peak concentration onthe 5th day after infection begins. Therefore, the virus can be easily detected in the samples from the nasopharynxup to seven days after the onset of the disease by PCR testing. In our country, real-time PCR-based analysis methodtargeting RNA-dependent RNA polymerase (RdRp) gene fragment is routinely used to diagnose Covid-19 disease.For the diagnosis of Covid-19; ideal samples for PCR-based analysis procedures are; swab samples taken from thenasopharynx and / or oropharynx and bronchoalveolar lavage fluid and sputum samples in patients with pneumonia.Besides, for the serological diagnose of Covid-19; ELISA kits that provide detection of spike proteins and recombinantnucleocapsid protein and rapid diagnostic kits have been developed to detect the antibodies or viral components innasopharyngeal secretions The World Health Organization (WHO) recommends the use of rapid antigen and antibodytests for the diagnosis of Covid-19 for surveillance studies.

Anahtar Kelimeler

COVID-19, Molecular, SARS-Cov-2, Serological

Kaynakça

• Bruning AHL, Lee ang MMG, Vos JMBW, Spijker R, de Jong MD,Wolthers KC, Pajkrt D. (2017). Rapid Tests for Influenza,Respiratory Syncytial Virus, and Other RespiratoryViruses: A Systematic Review and Meta-analysis. ClinInfect Dis. 2017: 65(6), 1026–32.
• CDC (2020).‘CDC 2019-Novel Coronavirus (2019-nCoV) Real-Time RT-PCR Diagnostic Panel.’ CDC-006-00019, Revision:02. Erişim Adresi: https://www.fda.gov/media/134922/download. Erişim tarihi: 28 Nisan 2020.
• European Commission (2020): ‘EU Recommendations fortesting strategies 2020. Erişim Adresi:https://ec.europa.eu/info/sites/info/files/covid19_eu_recommendations_on_testing_strategies_v2.pdfhttps://ec.europa.eu/info/sites/info/files/covid19_eu_recommendations_on_testing_strategies_v2.pdf Erişim tarihi: 28 Nisan 2020.
• Gorbalenya AE, Baker SC, Baric RS, Groot RJ, Drosten C,Gulyaeva AA, Haagmans BL, Lauber C, Leontovich AM,Neuman BW, Penzar D, Perlman S, Poon L, SamborskiyD, Sidorov IA, Solá GI, Ziebuhr J. (2020). The speciesSevere acute respiratory syndrome-related coronavirus:classifying 2019-nCoV and naming it SARS-CoV-2. NatMicrobiol. 5: 536–544. https://doi.org/10.1038/s41564-020-0695-z.
• Guo L, Ren L, Yang S, Xiao M, Chang D, Yang F, Dela CruzCS, Wang Y, Wu C, Xiao Y. (2020). Profiling Early HumoralResponse to Diagnose Novel Coronavirus Disease(COVID-19). Clin Infect Dis: ciaa310.
• Health BSoP. (2020). Serology testing for COVID-19’ JohnsHopkins Center for Health Security. Erişim Adresi: https://www.centerforhealthsecurity.org/resources/COVID-19/COVID-19-fact-sheets/200228-Serology-testing-COVID.pdf. Erişim tarihi: 28 Mart 2020.
• Hodinka RL. (2020). Point: is the era of viral culture over inthe clinical microbiology laboratory?’ J Clin Microbiol.2013:51(1):2-4.
• Lauer SA, Grantz KH, Bi Q, Jones FK, Zheng Q, Meredith HR,Azman AS, Reich NG, Lessler J. (2020). The IncubationPeriod of Coronavirus Disease 2019 (COVID-19) FromPublicly Reported Confirmed Cases: Estimation andApplication. Ann Intern Med, 172(9): 577-582.
• Li Z, Yi Y, Luo X, Xiong N, Liu Y, Li S. Sun R, Wang Y, Hu B, ChenW, Zhang Y, Wang J, Huang B, Lin Y, Yang J, Cai W, SunK, Pan W, Zhan Z, Chen L, YE F. (2020). Development andclinical application of a rapid IgM-IgG combined antibodytest for SARS-CoV-2 infection diagnosis. J Med Virol,10.1002/jmv.25727.
• Pan Y, Zhang D, Yang P, Poon LLM, Wang Q. (2020). Viral loadof SARS-CoV-2 in clinical samples. Lancet Infect Dis, 20:411–412.
• Park WB, Kwon NJ., Choi SJ, Kang CK, Choe PG, Kim JY, YunJ, Lee G-W, Seong MW, Kim NJ, Seo JS, Oh M. (2020).Virus Isolation from the First Patient with SARS-CoV-2 inKorea. J Korean Med Sci, 35(7):e84.
• Patel R, Babady E, Theel ES, Storch GA, Pinsky BA, St GeorgeK, Smith TC, Bertuzzi S. (2020). Report from the AmericanSociety for Microbiology COVID-19 International Summit23 March 2020: Value of Diagnostic Testing for SARSCoV-2/COVID-19. mBio, 26:11(2). pii: e00722-20.
• T.C. Sağlık Bakanlığı (2020). COVID-19 Rehberi. https://covid19bilgi.saglik.gov.tr/depo/rehberler/COVID-19_Rehberi.pdf Erişim tarihi: 28 Nisan 2020.
• Wang C, Horby P, Hayden FG, Gao F. (2020). A novel coronavirusoutbreak of global health concern. Lancet, 395:470–473.
• Wang W, Xu Y, Gao R, Lu R, Han K, Wu G, Tan W. (2020).Detection of SARS-CoV-2 in Different Types of ClinicalSpecimens. JAMA ,e203786.
• Winichakoon P, Chaiwarith R, Liwsrisakun C, Salee P, GoonnaA, Limsukon A. (2020). Negative nasopharyngeal andoropharyngeal swab does not rule out COVId-19. Journalof Clinical Microbiology, 58 (5): e00297-20.
• World Health Organization (WHO) (2020). Laboratory testing forcoronavirus disease 2019 (COVID-19) in suspected humancases: interim guidance. World Health Organization.https://apps.who.int/iris/handle/10665/331329 Erişimtarihi: 28 Nisan 2020.
• World Health Organization (2020).Advice on the use of pointof-care immunodiagnostic tests for COVID-19. https://www.who.int/news-room/commentaries/detail/adviceon-the-use-of-point-of-care-immunodiagnostic-testsfor-covid-19. Erişim tarihi: 28 Nisan 2020.
• Wölfel R, Corman VM, Guggemos W, Seilmaier M, Zange S,Müller MA (2020). Virological assessment of hospitalizedpatients with COVID-2019.Nature,10.1038/s41586-020-2196-x.
• Wu Y, Guo C, Tang L, Hong Z, Zhou J, Dong X. (2020). Prolongedpresence of SARS-CoV-2 viral RNA in faecal samples.Lancet Gastroenterol Hepatol. 5,434-435.
• Zhao J, Yuan Q, Wang H, Liu W, Liao X, Su Y. ve diğerleri (2020).Antibody responses to SARS-CoV-2 in patients of novelcoronavirus disease 2019, Clin Infect Dis., 28;ciaa344.
• Zou L, Ruan F, Huang M, Liang L, Huang H, Hong Z. ve diğerleri(2020). SARS-CoV-2 viral load in upper respiratoryspecimens of infected patients. N Engl J Med., 382:1177–1179.