Early and Effective Diagnosis of Sepsis Using Flow Cytometry

Abstract

Objective Sepsis is a major global health issue due to its high death and morbidity rates. To avoid the negative effects of sepsis and decrease mortality, it is vital to diagnose and treat it as soon as possible. Blood cultures can take up to 2 days to give result, and they are not always reliable. According to recent studies, neutrophil CD64 expression might be a sensitive and specific option for assessing sepsis. This study aimed to evaluate the diagnostic performance of a flow cytometry analysis for the expression of neutrophil CD64 in sepsis and its comparison with other standard tests in a tertiary care center.

Materials and Methods Prospective analysis on 40 blood samples from suspected sepsis patients admitted to intensive care units with criteria for the systemic inflammatory response syndrome on presentation was performed for expression of neutrophil CD64, C-reactive protein, procalcitonin, and complete blood count. Ten healthy volunteers were also enrolled in this prospective study. The laboratory results were compared in different groups.

Results The neutrophil CD64 had the highest diagnostic value to differentiate between patients of sepsis and nonsepsis groups with a sensitivity of 100% (95% confidence interval [CI]: 77.19–100%) and 100% (95% CI: 55.32–86.83%); specificity of 90.00% (95% CI: 59.58–99.49%) and 87.24% (95% CI: 66.69–99.61%); and likelihood ratio of 10.00 and 7.84, respectively.

Conclusion The neutrophil CD64 expression provides a more sensitive, specific, and novel marker for the early detection of sepsis in critically ill patients.

Keywords

CD64 - sepsis - biomarker - flow cytometry - intensive care unit

Ethical Approval

Approved by the Institutional Ethics Committee, reference no: 102ECM IIB/Thesis P-46, dated 05.09.2020.

Authors' Contribution

R.K. did study designing, conceptual analysis, data acquisition, and literature search. P.V. did data analysis and designing. A.S. did a literature search and proof correction. G.Y. did data analysis and proofreading. A.A. contributed to clinical studies and literature searches. H.D.R. contributed to clinical studies and design. S.P.V. did clinical studies and data analysis. U.S.S. did study design and literature search.

Publikationsverlauf

Artikel online veröffentlicht:
20. Oktober 2022

• References

• 1 Novosad SA, Sapiano MR, Grigg C. et al. Vital signs: epidemiology of sepsis: prevalence of health care factors and opportunities for prevention. MMWR Morb Mortal Wkly Rep 2016; 65 (33) 864-869
• 2 Vincent JL, Rello J, Marshall J. et al; EPIC II Group of Investigators. International study of the prevalence and outcomes of infection in intensive care units. JAMA 2009; 302 (21) 2323-2329
• 3 Rudd KE, Johnson SC, Agesa KM. et al. Global, regional, and national sepsis incidence and mortality, 1990-2017: analysis for the Global Burden of Disease Study. Lancet 2020; 395 (10219): 200-211
• 4 Kumar S, Tripathy S, Jyoti A, Singh SG. Recent advances in biosensors for diagnosis and detection of sepsis: a comprehensive review. Biosens Bioelectron 2019; 124-125: 205-215
• 5 Lever A, Mackenzie I. Sepsis: definition, epidemiology, and diagnosis. BMJ 2007; 335 (7625): 879-883
• 6 Sinha M, Jupe J, Mack H, Coleman TP, Lawrence SM, Fraley SI. Emerging technologies for molecular diagnosis of sepsis. Clin Microbiol Rev 2018; 31 (02) e00089
• 7 Groselj-Grenc M, Ihan A, Derganc M. Neutrophil and monocyte CD64 and CD163 expression in critically ill neonates and children with sepsis: comparison of fluorescence intensities and calculated indexes. Mediators Inflamm 2008; 2008: 202646
• 8 Cid J, Aguinaco R, Sánchez R, García-Pardo G, Llorente A. Neutrophil CD64 expression as marker of bacterial infection: a systematic review and meta-analysis. J Infect 2010; 60 (05) 313-319
• 9 Comstedt P, Storgaard M, Lassen AT. The systemic inflammatory response syndrome (SIRS) in acutely hospitalised medical patients: a cohort study. Scand J Trauma Resusc Emerg Med 2009; 17: 67
• 10 Annane D, Bellissant E, Cavaillon JM. Septic shock. Lancet 2005; 365 (9453): 63-78
• 11 Gerdes JS. Diagnosis and management of bacterial infections in the neonate. Pediatr Clin North Am 2004; 51 (04) 939-959 , viii–ix
• 12 Carvalho PR, Trotta EdeA. Avanços no diagnóstico e tratamento da sepse. [Advances in sepsis diagnosis and treatment] J Pediatr (Rio J) 2003; 79 (Suppl. 02) S195-S204
• 13 Migita K, Agematsu K, Yamazaki K. et al. Expression of CD64 on polymorphonuclear neutrophils in patients with familial Mediterranean fever. Clin Exp Immunol 2011; 164 (03) 365-372
• 14 Davis BH, Olsen SH, Ahmad E, Bigelow NC. Neutrophil CD64 is an improved indicator of infection or sepsis in emergency department patients. Arch Pathol Lab Med 2006; 130 (05) 654-661
• 15 Hsu KH, Chan MC, Wang JM, Lin LY, Wu CL. Comparison of Fcγ receptor expression on neutrophils with procalcitonin for the diagnosis of sepsis in critically ill patients. Respirology 2011; 16 (01) 152-160
• 16 Gros A, Roussel M, Sauvadet E. et al. The sensitivity of neutrophil CD64 expression as a biomarker of bacterial infection is low in critically ill patients. Intensive Care Med 2012; 38 (03) 445-452
• 17 Dal Ponte ST, Alegretti AP, Pilger DA. et al. Diagnostic accuracy of CD64 for sepsis in emergency department. J Glob Infect Dis 2018; 10 (02) 42-46
• 18 Icardi M, Erickson Y, Kilborn S, Stewart B, Grief B, Scharnweber G. CD64 index provides simple and predictive testing for detection and monitoring of sepsis and bacterial infection in hospital patients. J Clin Microbiol 2009; 47 (12) 3914-3919
• 19 Hoffmann JJ. Neutrophil CD64: a diagnostic marker for infection and sepsis. Clin Chem Lab Med 2009; 47 (08) 903-916
• 20 Lewis SM, Treacher DF, Bergmeier L. et al. Plasma from patients with sepsis up-regulates the expression of CD49d and CD64 on blood neutrophils. Am J Respir Cell Mol Biol 2009; 40 (06) 724-732
• 21 Zhou Y, Zhang Y, Johnson A, Venable A, Griswold J, Pappas D. Combined CD25, CD64, and CD69 biomarker panel for flow cytometry diagnosis of sepsis. Talanta 2019; 191: 216-221
• 22 Wang X, Li ZY, Zeng L. et al. Neutrophil CD64 expression as a diagnostic marker for sepsis in adult patients: a meta-analysis. [published correction appears in Critical Care. 2016;20(1):172] Crit Care 2015; 19 (01) 245
• 23 Dimoula A, Pradier O, Kassengera Z, Dalcomune D, Turkan H, Vincent JL. Serial determinations of neutrophil CD64 expression for the diagnosis and monitoring of sepsis in critically ill patients. Clin Infect Dis 2014; 58 (06) 820-829
• 24 Chauhan S, Hansa JA. Early diagnosis of sepsis through sepsis markers and sepsis index through flow cytometry technology. Asian J Pharm Clin Res 2017; 10: 145-148
• 25 Nargis W, Ibrahim M, Ahamed BU. Procalcitonin versus C-reactive protein: usefulness as biomarker of sepsis in ICU patient. Int J Crit Illn Inj Sci 2014; 4 (03) 195-199
• 26 Lãpez FR, Mote JD, FarÃas ON. Procalcitonin (PCT), C reactive protein (CRP) and its correlation with severity in early sepsis. Clin Rev Opinions 2011; 3 (03) 26-31
• 27 Ruiz-Alvarez MJ, García-Valdecasas S, De Pablo R. et al. Diagnostic efficacy and prognostic value of serum procalcitonin concentration in patients with suspected sepsis. J Intensive Care Med 2009; 24 (01) 63-71
• 28 Endo S, Aikawa N, Fujishima S. et al. Usefulness of procalcitonin serum level for the discrimination of severe sepsis from sepsis: a multicenter prospective study. J Infect Chemother 2008; 14 (03) 244-249
• 29 Pettilä V, Pentti J, Pettilä M, Takkunen O, Jousela I. Predictive value of antithrombin III and serum C-reactive protein concentration in critically ill patients with suspected sepsis. Crit Care Med 2002; 30 (02) 271-275