Evidence for SARS-CoV-2 Spike Protein in the Urine of COVID-19 Patients
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Abstract
Key Points
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Using an antigen capture assay to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike S1 protein, we found that the protein is present in the urine of 25% of patients with coronavirus disease 2019 (COVID-19).
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Further, we found that 24% and 21% of adult patients with COVID-19 have high levels of urine albumin and cystatin C, respectively.
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The presence of SARS-CoV-2 spike protein in the urine suggests renal abnormalities resulting from COVID-19.
Background
SARS-CoV-2 infection has, as of April 2021, affected >133 million people worldwide, causing >2.5 million deaths. Because the large majority of individuals infected with SARS-CoV-2 are asymptomatic, major concerns have been raised about possible long-term consequences of the infection.
Methods
Wedeveloped an antigen capture assay to detect SARS-CoV-2 spike protein in urine samples from patients with COVID-19whose diagnosis was confirmed by positive PCR results from nasopharyngeal swabs (NP-PCR+) forSARS-CoV-2. We used a collection of 233 urine samples from 132 participants from Yale New Haven Hospital and the Children’s Hospital of Philadelphia that were obtained during the pandemic (106 NP-PCR+ and 26 NP-PCR−), and a collection of 20 urine samples from 20 individuals collected before the pandemic.
Results
Our analysis identified 23 out of 91 (25%) NP-PCR+ adult participants with SARS-CoV-2 spike S1 protein in urine (Ur-S+). Interestingly, although all NP-PCR+ children were Ur-S−, one child who was NP-PCR− was found to be positive for spike protein in their urine. Of the 23 adults who were Ur-S+, only one individual showed detectable viral RNA in urine. Our analysis further showed that 24% and 21% of adults who were NP-PCR+ had high levels of albumin and cystatin C, respectively, in their urine. Among individuals with albuminuria (>0.3 mg/mg of creatinine), statistical correlation could be found between albumin and spike protein in urine.
Conclusions
Together, our data showed that one of four individuals infected with SARS-CoV-2 develop renal abnormalities, such as albuminuria. Awareness about the long-term effect of these findings is warranted.
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SciScore for 10.1101/2021.01.27.21250637: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Rabbit polyclonal anti-spike protein antibody purified using Protein G immunoaffinity chromatography was purchased from MyBioSource (MBS434243; 2µg/µL) and used as capture antibody in the capture ELISA. Rabbit polyclonal anti-spike protein antibodysuggested: Noneanti-spike proteinsuggested: NoneAfter 15-16 hours, the wells were washed four times with PBST, biotinylated polyclonal anti-COVID-19 antibodies (spike protein) were added at a concentration of 5µg/mL, and plates were incubated for … SciScore for 10.1101/2021.01.27.21250637: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Table 2: Resources
Antibodies Sentences Resources Rabbit polyclonal anti-spike protein antibody purified using Protein G immunoaffinity chromatography was purchased from MyBioSource (MBS434243; 2µg/µL) and used as capture antibody in the capture ELISA. Rabbit polyclonal anti-spike protein antibodysuggested: Noneanti-spike proteinsuggested: NoneAfter 15-16 hours, the wells were washed four times with PBST, biotinylated polyclonal anti-COVID-19 antibodies (spike protein) were added at a concentration of 5µg/mL, and plates were incubated for 1 hour at room temperature. anti-COVID-19suggested: NoneIn addition, to determine the specificity of polyclonal SARS-CoV-2 anti-spike antibodies (MyBioSource; MBS434243; 2µg/µL), different concentrations (0.1µg, 0.5µg and 1µg) of purified recombinant SARS-CoV-2 S1 (GenScript; Cat No Z03501) were probed using a Western blot (Figure 1E). anti-spikesuggested: NoneThe membrane was blocked with 5% milk (American Bio Cat No AB10109-00100) in PBST followed by treatment with polyclonal antibody against SARS-CoV-2 spike (S1+S2) protein raised in rabbit (MyBioSource MBS434243) and used at 1:1000 dilution in PBST S1+S2 ) proteinsuggested: NoneGoat anti-rabbit IgG horse radish peroxidase (HRP) conjugate (Thermofisher Scientific, USA, Cat No 31466) was used as the secondary antibody at 1:5000 dilution. anti-rabbit IgGsuggested: NoneResults from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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