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. 2021 Mar 23:8:655785.
doi: 10.3389/fmed.2021.655785. eCollection 2021.

Validation of a Quick Flow Cytometry-Based Assay for Acute Infection Based on CD64 and CD169 Expression. New Tools for Early Diagnosis in COVID-19 Pandemic

Alejandra Comins-Boo  1   2 , Maria Gutiérrez-Larrañaga  1   2 , Adriel Roa-Bautista  1   2 , Sandra Guiral Foz  1   2 , Mónica Renuncio García  1   2 , Elena González López  1   2 , Juan Irure Ventura  1   2 , María Carmen Fariñas-Álvarez  3   4 , David San Segundo  1   2 , Marcos López Hoyos  1   2
Affiliations

Validation of a Quick Flow Cytometry-Based Assay for Acute Infection Based on CD64 and CD169 Expression. New Tools for Early Diagnosis in COVID-19 Pandemic

Alejandra Comins-Boo et al. Front Med (Lausanne). .

Abstract

Objectives: Several parameters aid in deciphering between viral and bacterial infections; however, new tools should be investigated in order to reduce the time to results and proceed with an early target-therapy. Validation of a biomarker study, including CD64 and CD169 expression, was conducted. Material and Methods: Patients with active SARS-CoV-2 infection (ACov-2), bacterial infection (ABI), healthy controls, and antiretroviral-controlled chronic HIV infection were assessed. Whole blood was stained and, after lysing no-wash protocol, acquired by flow cytometry. The median fluorescence intensity (MFI) of CD64 and CD169 was measured in granulocytes, monocytes, and lymphocytes. The CD64 MFI ratio granulocytes to lymphocytes (CD64N) and CD169 MFI ratio monocytes to lymphocytes (CD169Mo) were evaluated as biomarkers of acute bacterial and viral infection, respectively. Results: A CD64N ratio higher than 3.3 identified patients with ABI with 83.3 and 85.9% sensitivity and specificity, with an area under the curve (AUC) of 83.5%. In contrast, other analytic or hematological parameters used in the clinic had lower AUC compared with the CD64N ratio. Moreover, a CD169Mo ratio higher than 3.3 was able to identify ACov-2 with 91.7 and 89.8 sensitivity and specificity, with the highest AUC (92.0%). Conclusion: This work confirms the previous data of CD64N and CD169Mo ratios in an independent cohort, including controlled chronic viral HIV infection patients as biomarkers of acute bacterial and viral infections, respectively. Such an approach would benefit from quick pathogen identification for a direct-therapy with a clear application in different Health Care Units, especially during this COVID pandemic.

Keywords: CD169; CD64; SARS-CoV-2; biomarkers; flow cytometry; validation.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Biomarkers of acute infection. Representative histograms of the median fluorescence intensity in neutrophil granulocytes (purple), lymphocytes (blue), and monocytes (green) of CD64 (A) and CD169 (B) expression in healthy control (HC), acute bacterial infection (ABI), and acute SARS-CoV-2 infection (ACov2) patient to calculate the ratio CD64 and CD169 as described in Material and Methods section. The ratios of CD64N (A) and CD169Mo are depicted (B). The ratio CD64N (C) and CD169Mo (D) in HC (red light circles), ABI (green squares), chronically HIV infected patients (HIV, blue triangles), and acute SARS-CoV-2 (ACov-2, yellow triangles) are depicted. The dotted lines represent the cut-off value for calculating the Receiver Operational Characteristic (ROC) curve. The ROC curve of different parameters used to decipher acute infections are shown, C-reactive protein (blue line), neutrophil frequency (red line), absolute neutrophil count (green line), lymphocyte frequency (yellow line), lymphocyte count (gray line), CD64N ratio (orange line) (E), and CD169Mo ratio (orange line) (F). ***p <0.001 and *p < 0.05.
Figure 2
Figure 2
Scatter plot showing the correlation between CD169Mo ratio and time in days from positive PCR of SARS-CoV-2.
See this image and copyright information in PMC

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