Abstract
We have applied nuclear magnetic resonance spectroscopy based plasma phenotyping to reveal diagnostic molecular signatures of SARS-CoV-2 infection via combined diffusional and relaxation editing (DIRE). We compared plasma from healthy age-matched controls ( n = 26) with SARS-CoV-2 negative non-hospitalized respiratory patients and hospitalized respiratory patients ( n = 23 and 11 respectively) with SARS-CoV-2 rRT-PCR positive respiratory patients ( n = 17, with longitudinal sampling time-points). DIRE data were modelled using principal component analysis and orthogonal projections to latent structures discriminant analysis (O-PLS-DA), with statistical cross-validation indices indicating excellent model generalization for the classification of SARS-CoV-2 positivity for all comparator groups (area under the receiver operator characteristic curve = 1). DIRE spectra show biomarker signal combinations conferred by differential concentrations of metabolites with selected molecular mobility properties. These comprise the following: (a) composite N -acetyl signals from α-1-acid glycoprotein and other glycoproteins (designated GlycA and GlycB) that were elevated in SARS-CoV-2 positive patients [ p = 2.52 × 10 -10 (GlycA) and 1.25 × 10 -9 (GlycB) vs controls], (b) two diagnostic supramolecular phospholipid composite signals that were identified (SPC-A and SPC-B) from the – + N-(CH 3 ) 3 choline headgroups of lysophosphatidylcholines carried on plasma glycoproteins and from phospholipids in high-density lipoprotein subfractions (SPC-A) together with a phospholipid component of low-density lipoprotein (SPC-B). The integrals of the summed SPC signals (SPC total ) were reduced in SARS-CoV-2 positive patients relative to both controls ( p = 1.40 × 10 -7 ) and SARS-CoV-2 negative patients ( p = 4.52 × 10 -8 ) but were not significantly different between controls and SARS-CoV-2 negative patients. The identity of the SPC signal components was determined using one and two dimensional diffusional, relaxation, and statistical spectroscopic experiments. The SPC total /GlycA ratios were also significantly different for control versus SARS-CoV-2 positive patients ( p = 1.23 × 10 -10 ) and for SARS-CoV-2 negatives versus positives ( p = 1.60 × 10 -9 ). Thus, plasma SPC total and SPC total /GlycA are proposed as sensitive molecular markers for SARS-CoV-2 positivity that could effectively augment current COVID-19 diagnostics and may have value in functional assessment of the disease recovery process in patients with long-term symptoms.
【초록키워드】 COVID-19, Structure, SARS-CoV-2, Biomarker, Hospitalized, SARS-COV-2 infection, diagnostic, classification, Symptoms, diagnostics, principal component analysis, rRT-PCR, Patient, Control, plasma, glycoprotein, metabolites, molecular, Glycoproteins, respiratory, nuclear magnetic resonance, patients, Magnetic resonance spectroscopy, metabolite, marker, Phospholipids, High-density lipoprotein, Low-density lipoprotein, Combination, Concentration, Analysis, Relaxation, Functional assessment, molecular markers, Principal component, discriminant analysis, choline, negative patients, identity, following, SARS-CoV-2 positivity, Comparator group, low-density, component, positive, SARS-CoV-2 positive patients, phospholipid, nuclear, high-density, statistical, Receiver operator characteristic, selected, carried, significantly, healthy, the disease, elevated, applied, reduced, functional, was determined, experiments, age-matched control, not significantly different, SARS-CoV-2 positive patient, 【제목키워드】 SARS-CoV-2, NMR, Spectroscopy, signature, proton,