Аннотация
В декабре 2019 года в Китае была зарегистрирована новая форма коронавируса (SARS-CoV-2), вызвавшая к настоящему времени более 690 млн случаев заражения и более 6,8 млн смертей от коронавирусной болезни (COVID-19) по всему миру. В связи с этим крайне актуальным является изучение маркеров раннего прогноза тяжести инфекции COVID-19 у пациентов. Цель исследования — проанализировать клинико-лабораторные показатели у пациентов в зависимости от инфицированности штаммами SARS-CoV-2 «Ухань», «Альфа» и «Дельта» SARS-CoV-2 в северо-западном регионе Российской Федерации.
Материал и методы. В ретроспективное когортное исследование включены клинико-биохимические данные, полученные от 6 780 пациентов (2 932 мужчин и 3 848 женщин в возрасте 51,2±11,6 лет) с положительными результатами ПЦР тестов на наличие РНК SARS-CoV-2, проходивших лечение в Городской больнице № 40 Санкт-Петербурга с 01.09.2020 по 01.08.2021 года. Пациенты были разделены на 3 группы в зависимости от инфицированности штаммами коронавируса: «Ухань» (n=1276), «Альфа» (n=2016) и «Дельта» (n=3398). Всем пациентам выполнялся клинический анализ крови с оценкой активности лактатдегидрогеназы (ЛДГ), аланинаминотрансферазы (АЛТ), аспартатаминотрансферазы (АСТ), уровня С реактивного белка (СРБ), интерлейкина-6 (ИЛ-6), общего билирубина, креатинина, общего белка, ферритина в сыворотке крови; активированного парциального тромбопластинового времени (АПТВ), фибриногена, D-димера в плазме крови; уровня гемоглобина, количества эритроцитов, тромбоцитов, лейкоцитов, лимфоцитов, нейтрофилов, эозинофилов в цельной крови; скорости клубочковой фильтрации (СКФ), а также проводилась компьютерная томография (КТ) органов грудной клетки.
Результаты. У пациентов (n=3 398), инфицированных штаммом «Дельта» SARS-CoV-2 при поступлении достоверно чаще повышались уровни АСТ, АЛТ, ЛДГ, ферритина, фибриногена по сравнению с пациентами, инфицированными штаммами «Ухань» и «Альфа». У больных группы «Дельта» чаще снижалось количество эозинофилов, эритроцитов, уровень общего белка, билирубина и гемоглобина чем у пациентов групп «Ухань» и «Альфа». У пациентов групп «Альфа» и «Дельта» наблюдалось статистически значимое повышение уровней АЛТ (на 3-и и 6-е сутки), АПТВ (на 3-и и 6-е сутки), АСТ (на 3, 6-е сутки), D-димера (на 12-е сутки), ИЛ-6 (на 3, 9, 12-е сутки), ЛДГ (3, 12-е сутки), СРБ (на 3, 6, 9, 12-е сутки), тромбоцитов (3, 6-е сутки), ферритина (на 3-и сутки), эозинофилов (на 3, 6, 12-е сутки) по сравнению с пациентами группы «Ухань» (n=1276). Наиболее значимыми предикторами летального исхода в группах «Ухань» являются: максимальный уровень ИЛ-6, СРБ, креатинина, лейкоцитов, степень снижения СКФ; в группе «Альфа»: уровень общего билирубина, ИЛ-6, СРБ, выраженность лимфоцитопении и падения СКФ и в группе «Дельта»: максимальный уровень ИЛ-6, СРБ, общего билирубина, степень тромбоцитопении и снижения СКФ.
Заключение. Высокие уровни лимфоцитов у пациентов группы «Ухань» в начальные временные точки по сравнению с группами «Альфа» и «Дельта» указывают на сильную активацию иммунной системы организма в ответ на вирусную инфекцию. Панцитопения, снижение уровней общего белка и повышение СРБ, ИЛ-6, ЛДГ у пациентов групп «Альфа» и, особенно, «Дельта», свидетельствует о более сильной вирулентности вируса по сравнению со штаммом «Ухань». Оценка в динамике уровней этих аналитов может быть использована для прогнозирования летального исхода от COVID-19 у пациентов.
Annotation
In December 2019, a new form of coronavirus (SARS-CoV-2) was recorded in China, which caused more than 690 million cases of infection and more than 6.8 million deaths from the coronavirus disease (COVID-19). In this regard, it extremely urgent to study markers for predicting the severity of a COVID-19 infection. The aim of the study was to analyze clinical and laboratory parameters depending on the infection of patients with Wuhan, Alpha and Delta strains of SARS-CoV-2 in the northwestern region of the Russian Federation.
Material and methods. The retrospective cohort study included clinical and biochemical data obtained from 6780 patients (2,932 men and 3,848 women aged 51.2±11.6) who showed positive PCR test results for the presence of SARS-CoV-2 RNA (using a PCR test), who were treated at the City Hospital No. 40 of Saint-Petersburg from 09/01/2020 to 10/15/2021. Patients were divided into 3 groups depending on infection with coronavirus strains: «Wuhan» (n=1276), «Alpha» (n=2016) and «Delta» (n=3398). A detailed clinical blood analysis with activity score of lactate dehydrogenase (LDH), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and the levels of C-reactive protein (CRP), interleukin-6 (IL-6), total bilirubin, creatinine, total protein, ferritin in the blood serum; activated partial thromboplastin time (APTT), fibrinogen, D-dimer in the blood plasma; the levels of hemoglobin, amounts of erythrocytes, platelets, leukocytes, lymphocytes, neutrophils, eosinophils in the whole blood; glomerular filtration rate (GFR) were performed in all patients, as well as computed tomography (CT) of the chest.
Results. In patients (n=3398) infected with the Delta strain of SARS-CoV-2 on the admission, the levels of AST, ALT, LDH, ferritin, fibrinogen were significantly more likely to increase compared to patients infected with the Wuhan and Alpha strains. In patients of the «Delta» group, the number of eosinophils, erythrocytes, the level of total protein, bilirubin and hemoglobin often decreases than in patients of the «Wuhan» and «Alpha» groups. In patients of the «Alpha» and «Delta» groups, there was a statistically significant increase in the levels of ALT (on days 3, 6), APTT (on days 3, 6), AST (on days 3, 6), D-dimer (on 3-12 days), IL-6 (on days 3, 9, 12), LDH (on days 3, 12), CRP (on days 3-12), platelets (on days 3, 6), ferritin (on day 3), eosinophils (on days 3, 6, 12) compared with patients of the Wuhan group (n=1276). The most significant predictors of death in the Wuhan groups are: the maximum level of IL-6, CRP, creatinine, leukocytes, decline in GFR rate, Alpha group: the level of total bilirubin, IL-6, CRP, severity of lymphocytopenia and decline in GFR, and in the «Delta» group: the maximum level of IL-6, CRP, total bilirubin, the thrombocytopenia rate and decline in GFR.
Conclusion. High levels of lymphocytes in Wuhan patients at initial time points compared to Alpha and Delta groups indicate a strong activation of the body’s immune system in response to a viral infection. Pancytopenia, a decrease in total protein levels and an increase in CRP, IL-6, LDH in patients of the Alpha and, especially, Delta groups, indicates a stronger virulence of the virus and its immunosuppressive effect on the body compared to the Wuhan strain. Evaluation of the levels of these analytes over time can be used to predict death from COVID-19 in the patients.
Key words: COVID-19; biochemical laboratory analytes; severity; COVID-19 severity prediction; Wuhan; Alpha and Delta strains of SARS-CoV-2.
Список литературы
1.Freund N.T., Roitburd-Berman A., Sui J., Marasco W.A., Gershoni J.M. Reconstitution of the receptor-binding motif of the SARS coronavirus. Protein Eng. Des. Sel. 2015; 28(12): 567‒75. DOI: 10.1093/protein/gzv052.
2.Mackay I.M., Arden K.E. MERS coronavirus: diagnostics, epidemiology and transmission. Virol J. 2015; 12: 222. DOI: 10.1186/s12985-015-0439-5.
3.COVID-19 coronavirus pandemic. Available at: https://www. worldomыeters.info/coronavirus/#countries
4.Interim guidelines: prevention, diagnosis and treatment of novel coronavirus infection (COVID-19). Version 15 [Vremennye metodicheskie rekomendatsii: profilaktika, diagnostika I lechenie novoy koronavirusnoy infektsii (COVID-19). Versiya 15]. Moscow: Ministry of Health of the Russian Federation; 2022. (in Russian)
5.Carrazco-Montalvo A., Bruno A., de Mora D., Olmedo M., Garces J., Paez M. et al. First Report of SARS-CoV-2 Lineage B.1.1.7 (Alpha Variant) in Ecuador, January 2021. Infect. Drug Resist. 2021; 14: 5183–8. Doi: 10.2147/IDR.S319439.
6.World Health Organization. SARS-CoV-2 Variants. Available at: https://www.who.int/westernpacific/emergencies/covid-19/information/covid-19-variants.
7.Sabino E.C., Buss L.F., Carvalho M.P.S., Prete C.A. Jr., Crispim M.A.E., Fraiji N.A. et al. Resurgence of COVID-19 in Manaus, Brazil, despite high seroprevalence. Lancet. 2021; 397: 452–5. DOI:10.1016/S0140-6736(21)00183-5.
8.Shiehzadegan S., Alaghemand N., Fox M., Venketaraman V. Analysis of the Delta Variant B.1.617.2 COVID-19. Clin. Pract.2021; 11: 778–84. DOI:10.3390/clinpract11040093.
9.Sharma V., Rai H., Gautam D.N.S., Prajapati P.K., Sharma R. Emerging evidence on Omicron (B.1.1.529) SARS-CoV-2 variant. J. Med. Virol. 2022; 94: 1876–85. DOI:10.1002/jmv.27626.
10.O’Toole Á., Scher E., Underwood A., Jackson B., Hill V., McCrone J.T. et al. Assignment of epidemiological lineages in an emerging pandemic using the pangolin tool. Virus Evol. 2021; 7(2) :veab064. DOI: 10.1093/ve/veab064.
11.La Rosa G., Brandtner D., Mancini P., Veneri C., , Ferraro G. B., Bonadonna L. et al. Key SARS-CoV-2 Mutations of Alpha, Gamma, and Eta Variants Detected in Urban Wastewaters in Italy by Long-Read Amplicon Sequencing Based on Nanopore Technology. Water. 2021; 13: 2503. DOI:10.3390/w13182503.
12.Yang X., Yu Y., Xu J., Shu H., Xia J., Liu H. et al. Clinical course and outcomes of critically ill patients with SARS-CoV-2 pneumonia in Wuhan, China: A single-centered, retrospective, observational study. Lancet Respir. Med. 2020; 8: 475–81. DOI:10.1016/S2213-2600(20)30079-5.
13.Shitao R.A.O., Lau A., So, H.-C. Exploring diseases/traits and blood proteins causally related to expression of ACE2, the putative receptor of 2019-nCov: A mendelian randomization analysis. Diabetes Care. 2020; 43: 1416–26. DOI:10.2337/dc20-0643.
14.Fang L., Karakiulakis G., Roth M. Are patients with hypertension and diabetes mellitus at increased risk for COVID-19 infection? Lancet Respir. Med. 2020; 8: e21. DOI:10.1016/S2213-2600(20)30116-8.
15.Wiersinga W.J., Rhodes A., Cheng A.C., Peacock S.J., Prescott H.C. Pathophysiology, transmission, diagnosis, and treatment of Coronavirus Disease 2019 (COVID-19): A review. JAMA. 2020; 324: 782–93. DOI:10.1001/jama.2020.12839.
16.Alzaabi A.H., Ahmed L.A., Rabooy A.E., Zaabi A.A., Alkaabi M., AlMahmoud F. et al. Longitudinal changes in IgG levels among COVID-19 recovered patients: A prospective cohort study. PLoS One. 2021; 16: e0251159. DOI:10.1371/journal.pone.0251159.
17.Gallais F., Velay A., Nazon C., Wendling M.-J., Partisani M., Sibilia J. et al. Intrafamilial exposure to SARS-CoV-2 induces cellular immune response without seroconversion. Emerg. Infect. Dis. 2021; 27: 113–21. DOI:10.3201/eid2701.203611.
18.Chen G., Wu D., Guo W., Cao Y., Huang D., Wang H. et al. Clinical and immunological features of severe and moderate coronavirus disease 2019. J. Clin. Investig. 2020; 130: 2620–9. DOI:10.1172/JCI137244.
19.Bhakta S., Sanghavi D.K., Johnson P.W., Kunze K.L., Neville M.R., Wadei H.M. et al. Clinical and laboratory profiles of the SARS-CoV-2 Delta variant compared with pre-Delta variants.. Int. J. Infect. Dis 2022; 120:88-95. DOI: 10.1016/j.ijid.2022.04.050.
20.Ward T., Glaser A., Overton C.E., Carpenter .B, Gent N., Seale A.C. Replacement dynamics and the pathogenesis of the Alpha, Delta and Omicron variants of SARS-CoV-2. Epidemiol Infect 2022; 151:e32. DOI: 10.1017/S0950268822001935.
21.Meletis G., Tychala A., Meletis N.G., Tychala A., Ntritsos G., Verrou E. et al. Variant-Related Differences in Laboratory Biomarkers among Patients Affected with Alpha, Delta and Omicron: A Retrospective Whole Viral Genome Sequencing and Hospital-Setting Cohort Study. Biomedicines. 2023; 11: 1143. DOI: 10.3390/biomedicines11041143.
22.Timur D., Demirpek U., Ertek E., Aydın Ö.Ç., Karabıyık T., Kayadibi H. Comparasion of SARS-COV-2 Wuhan and Alpha variants: clinical and laboratory highlights. Int. J. Med. Biochem. 2022;5(3):132-136. DOI: 10.14744/ijmb.2022.69772.
23.Gushchin V.A., Pochtovyi A.A., Kustova D.D., Ogarkova D.A., Tarnovetskii I.Y., Belyaeva E.D. et al. Dynamics of SARS-CoV-2 Major Genetic Lineages in Moscow in the Context of Vaccine Prophylaxis. Int. J. Mol. Sci 2022; 23: 14670. DOI: 10.3390/ijms232314670.
24.National Early Warning Score (NEWS) 2: NEWS2 and deterioration in COVID-19. London: RCP, 2017. Available at: https://www. rcplondon.ac.uk/news/news2-and-deterioration-covid-19.
25.Inui S., Fujikawa A, Jitsu M, Kunishima N, Watanabe S, Suzuki Y. et al. Chest CT Findings in Cases from the Cruise Ship Diamond Princess with Coronavirus Disease (COVID-19). Radiol Cardiothorac Imaging. 2020; 2(2): e200110. DOI: 10.1148/ryct.2020200110.
26.Laboratory diagnosis of COVID-19. Guidelines MP 3.1.0169-20 (as amended by MP 3.1.0174-20 “Changes No. 1 to MP 3.1.0170-20 “Laboratory diagnostics of COVID-19”, approved by Rospotrebnadzor on 30.04.2020). [Laboratornaya diagnostika COVID-19. Metodicheskie rekomendatsii MR 3.1.0169-20]. Moscow; 2020. (in Russian)
27.Shcherbak S. G., Anisenkova A. Yu., Mosenko S.V., Glotov O. S., Chernov A.N., Apalko S. V. et al. Basic predictive risk factors for cytokine storms in COVID-19 patients. Front Immunol. 2021; 12: 745515. DOI: 10.3389/fimmu.2021.745515.
28.Automated Hematology Analyzer XN series (Для системы XN-1000); Japan: Sysmex Corporation; 2013.
29.Reagent STA® — Cephascreen®; France: Diagnostica Stago S.A.S; 2017.
30.COBAS Elecsys IL-6; USA: Roche Diagnostics GmbH; 2020.
31.Herold T., Jurinovic V., Arnreich C., Lipworth B. J., Hellmuth J. C., von Bergwelt-Baildon M. et al. Elevated levels of interleukin-6 and CRP predict the need for mechanical ventilation in COVID-19. J. Allergy Clin. Immunol. 2020;146(1):128–136.e4. DOI: 10.1016/j. jaci.2020.05.008.
32.BRAHMS PCT-Q Immunochromatographic test for the detection of PCT (procalcitonin) in human serum and plasma, Germany; 2007.
33.ASPARTATE AMINOTRANSFERASE in ARCHITECT cSystems™ and AEROSET; USA: Abbott Laboratories; 2009.
34.C-reactive protein in ARCHITECT c Systems™ and AEROSET, USA: Abbott Laboratories; 2008.
35.CREATININE in ARCHITECT cSystems™ and AEROSET; USA: Abbott Laboratories; 2006.
36.LACTATE DEHYDROGENASE in ARCHITECT cSystems™ and AEROSET; USA: Abbott Laboratories; 2009.
37.The ARCHITECT Ferritin assay is a Chemiluminescent Microparticle Immunoassay (CMIA) for the quantitative determination of ferritin in human serum and plasma ARCHITECT cSystems™ and AEROSET; USA: Abbott Laboratories; 2010.
38.STA® — Liatest ® D-Di kit (REF 00515) Immuno-Turbidimetric Assay of D-Dimer; France: Diagnostica Stago S.A.S; 2017.
39.Thermo Scientific GeneJET RNA Purification Kit #K0731, #K0732. Available at: http://tools.thermofisher.com/content/sfs/manuals/MAN0012664_GeneJET_RNA_Purification_UG.pdf.
40.Agilent 4200 TapeStation System. System Manual. Germany: Agilent Technologies; 2020.
41.Mint-2 cDNA synthesis kit Cat # SK005. User manual. M.: Evrogen; 2020.
42.Thermo Scientific Maxima H Minus Double-Stranded cDNA Synthesis Kit #K2561. Thermo Fisher Scientific Inc.; 2013.
43.GeneFinder™ COVID-19 PLUS RealAmp Kit. Instructions for Use. Korea: OSANG Healthcare Co., Ltd.; 2021.
44.van Belle G., Fisher L. D., Heagerty P. J., Lumley T. Biostatistics : a methodology for the helth sciences; eds: Fisher L. D., van Belle G. Canada: Jonh Wiley and Sons Inc.; 2004.
45.Medical statistics. https://medstatistic.ru/calculators/calcodds.html.
46.Urazov S., Chernov A., Popov O., Klenkova N., Sushentseva N., Polkovnikova I. et al. Secretory phospholipase A2 and interleukin-6 levels as predictive markers of the severity and outcome of the patients with COVID-19 infections. Int. J. of Mol. Sci. 2023; 24: 5540. DOI:10.3390/ijms24065540.
47.Liu Z., Li J., Chen D., Gao R., Zeng W., Chen S. et al. Dynamic interleukin-6 level changes as a prognostic indicator in patients with COVID-19. Front Pharmacol. 2020; 11:1093. DOI: 10.3389/fphar.2020.01093.
48.Grifoni E., Valoriani A., Cei F., Lamanna R., Grazia Gelli A. M., Ciambotti B. et al. Interleukin-6 as prognosticator in patients with COVID-19. J. Infect. 2020;81 (3):452–82. DOI: 10.1016/j. jinf.2020.06.008.
49.Nahari A.D., Son M.B.F., Newburger J.W., Reis B. Y. An integrated framework for identifying clinical-laboratory indicators for novel pandemics: COVID-19 and MIS-C. npj Digit. Med. 2022; 5: 9. DOI:10.1038/s41746-021-00547-9.
50.Boytsov S.A., Pogosova N.V., Paleev F.N., Ezhov M.V., Komarov A.L., Pevsner D.V., Gruzdev K.A., Barinova I.V., Suvorov A.Yu.,Alekseeva I.A., Milko O.V. Clinical Characteristics and Factors Associated with Poor Outcomes in Hospitalized Patients with Novel Coronavirus Infection COVID-19. Kardiologiya. 2021;61(2):4-14. DOI:10.18087/cardio.2021.2.n1532. (in Russian)
51.Sharifpour M., Rangaraju S, Liu M, Alabyad D, Nahab FB, Creel-Bulos CM. et al. C-Reactive protein as a prognostic indicator in hospitalized patients with COVID-19. PLoS One. 2020; 15(11) :e0242400. DOI: 10.1371/journal.pone.0242400.
52.Wang Q., Cheng J., Shang J., Wang Y., Wan J., Yan Y.Q. et al. Clinical value of laboratory indicators for predicting disease progression and death in patients with COVID-19: a retrospective cohort study. BMJ Open. 2021;11(10): e043790. DOI:10.1136/bmjopen-2020-043790.
53.Huang H., Cai S., Li Y., Li Y., Fan Y., Li L. et al. Prognostic factors for COVID-19 pneumonia progression to severe symptom based on the earlier clinical features: a retrospective analysis. Front Med (Lausanne). 2020; 7: 557453. DOI:10.3389/fmed.2020.557453.
54.Guo Y., Liu Y., Lu J., Fan R., Zhang F., Yin X. et al. Development and validation of an early warning score (EWAS) for predicting clinical deterioration in patients with coronavirus disease 2019. MedRxiv. 2020. DOI:10.1101/2020.04.17.20064691.