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      Alterations in the human oral and gut microbiomes and lipidomics in COVID-19

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          Abstract

          Objective

          To characterise the oral microbiome, gut microbiome and serum lipid profiles in patients with active COVID-19 and recovered patients; evaluate the potential of the microbiome as a non-invasive biomarker for COVID-19; and explore correlations between the microbiome and lipid profile.

          Design

          We collected and sequenced 392 tongue-coating samples, 172 faecal samples and 155 serum samples from Central China and East China. We characterised microbiome and lipid molecules, constructed microbial classifiers in discovery cohort and verified their diagnostic potential in 74 confirmed patients (CPs) from East China and 37 suspected patients (SPs) with IgG positivity.

          Results

          Oral and faecal microbial diversity was significantly decreased in CPs versus healthy controls (HCs). Compared with HCs, butyric acid-producing bacteria were decreased and lipopolysaccharide-producing bacteria were increased in CPs in oral cavity. The classifiers based on 8 optimal oral microbial markers (7 faecal microbial markers) achieved good diagnostic efficiency in different cohorts. Importantly, diagnostic efficacy reached 87.24% in the cross-regional cohort. Moreover, the classifiers successfully diagnosed SPs with IgG antibody positivity as CPs, and diagnostic efficacy reached 92.11% (98.01% of faecal microbiome). Compared with CPs, 47 lipid molecules, including sphingomyelin (SM)(d40:4), SM(d38:5) and monoglyceride(33:5), were depleted, and 122 lipid molecules, including phosphatidylcholine(36:4p), phosphatidylethanolamine (PE)(16:0p/20:5) and diglyceride(20:1/18:2), were enriched in confirmed patients recovery.

          Conclusion

          This study is the first to characterise the oral microbiome in COVID-19, and oral microbiomes and lipid alterations in recovered patients, to explore their correlations and to report the successful establishment and validation of a diagnostic model for COVID-19.

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          Most cited references36

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          SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor

          Summary The recent emergence of the novel, pathogenic SARS-coronavirus 2 (SARS-CoV-2) in China and its rapid national and international spread pose a global health emergency. Cell entry of coronaviruses depends on binding of the viral spike (S) proteins to cellular receptors and on S protein priming by host cell proteases. Unravelling which cellular factors are used by SARS-CoV-2 for entry might provide insights into viral transmission and reveal therapeutic targets. Here, we demonstrate that SARS-CoV-2 uses the SARS-CoV receptor ACE2 for entry and the serine protease TMPRSS2 for S protein priming. A TMPRSS2 inhibitor approved for clinical use blocked entry and might constitute a treatment option. Finally, we show that the sera from convalescent SARS patients cross-neutralized SARS-2-S-driven entry. Our results reveal important commonalities between SARS-CoV-2 and SARS-CoV infection and identify a potential target for antiviral intervention.
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            UPARSE: highly accurate OTU sequences from microbial amplicon reads.

            Amplified marker-gene sequences can be used to understand microbial community structure, but they suffer from a high level of sequencing and amplification artifacts. The UPARSE pipeline reports operational taxonomic unit (OTU) sequences with ≤1% incorrect bases in artificial microbial community tests, compared with >3% incorrect bases commonly reported by other methods. The improved accuracy results in far fewer OTUs, consistently closer to the expected number of species in a community.
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              Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy.

              The Ribosomal Database Project (RDP) Classifier, a naïve Bayesian classifier, can rapidly and accurately classify bacterial 16S rRNA sequences into the new higher-order taxonomy proposed in Bergey's Taxonomic Outline of the Prokaryotes (2nd ed., release 5.0, Springer-Verlag, New York, NY, 2004). It provides taxonomic assignments from domain to genus, with confidence estimates for each assignment. The majority of classifications (98%) were of high estimated confidence (> or = 95%) and high accuracy (98%). In addition to being tested with the corpus of 5,014 type strain sequences from Bergey's outline, the RDP Classifier was tested with a corpus of 23,095 rRNA sequences as assigned by the NCBI into their alternative higher-order taxonomy. The results from leave-one-out testing on both corpora show that the overall accuracies at all levels of confidence for near-full-length and 400-base segments were 89% or above down to the genus level, and the majority of the classification errors appear to be due to anomalies in the current taxonomies. For shorter rRNA segments, such as those that might be generated by pyrosequencing, the error rate varied greatly over the length of the 16S rRNA gene, with segments around the V2 and V4 variable regions giving the lowest error rates. The RDP Classifier is suitable both for the analysis of single rRNA sequences and for the analysis of libraries of thousands of sequences. Another related tool, RDP Library Compare, was developed to facilitate microbial-community comparison based on 16S rRNA gene sequence libraries. It combines the RDP Classifier with a statistical test to flag taxa differentially represented between samples. The RDP Classifier and RDP Library Compare are available online at http://rdp.cme.msu.edu/.
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                Author and article information

                Journal
                Gut
                Gut
                gutjnl
                gut
                Gut
                BMJ Publishing Group (BMA House, Tavistock Square, London, WC1H 9JR )
                0017-5749
                1468-3288
                March 2021
                31 March 2021
                : gutjnl-2020-323826
                Affiliations
                [1 ]departmentDepartment of Infectious Diseases , the First Affiliated Hospital of Zhengzhou University , Zhengzhou, Henan, China
                [2 ]departmentGene Hospital of Henan Province , the First Affiliated Hospital of Zhengzhou University , Zhengzhou, Henan, China
                [3 ]departmentState Key Laboratory for Diagnosis and Treatment of Infectious Disease , the First Affiliated Hospital, School of Medicine, Zhejiang University , Hangzhou, Zhejiang, China
                [4 ]departmentDepartment of Clinical Laboratory , Henan Provincial Chest Hospital , Zhengzhou, Henan, China
                [5 ]departmentDepartment of General Surgery , Guangshan County People’s Hospital , Xinyang, Henan, China
                [6 ]departmentDepartment of Hepatobiliary and Pancreatic Surgery , the First Affiliated Hospital, School of Medicine, Zhejiang University , Hangzhou, Zhejiang, China
                [7 ]Shanghai Mobio Biomedical Technology Co, Ltd , Shanghai, Shanghai, China
                [8 ]departmentClinical Laboratory Diagnostics, Medical Technology College , Beihua University , Jilin, Jilin, China
                [9 ]departmentInternational Peace Maternity and Child Health Hospital , School of Medicine, Shanghai Jiao Tong University , Shanghai, Shanghai, China
                [10 ]departmentDepartment of Oncology , the First Affiliated Hospital of Zhengzhou University , Zhengzhou, Henan, China
                [11 ]departmentShulan (Hangzhou) Hospital , Zhejiang Shuren University Shulan International Medical College , Hangzhou, Zhejiang, China
                Author notes
                [Correspondence to ] Dr Zhigang Ren, Department of Infectious Diseases, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China; fccrenzg@ 123456zzu.edu.cn ; Dr Zujiang Yu, Department of Infectious Diseases, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China; johnyuem@ 123456zzu.edu.cn ; Professor Lanjuan Li, State Key Laboratory for Diagnosis and Treatment of Infectious Disease, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China; ljli@ 123456zju.edu.cn
                Author information
                http://orcid.org/0000-0003-0798-3444
                http://orcid.org/0000-0002-3273-1268
                http://orcid.org/0000-0002-3150-5269
                http://orcid.org/0000-0002-2998-4918
                http://orcid.org/0000-0001-6945-0593
                Article
                gutjnl-2020-323826
                10.1136/gutjnl-2020-323826
                8042598
                33789966
                4534d7d8-6b30-4845-a056-976cd02329d7
                © Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

                This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See:  http://creativecommons.org/licenses/by-nc/4.0/.

                History
                : 11 December 2020
                : 04 February 2021
                : 17 February 2021
                Funding
                Funded by: http://dx.doi.org/10.13039/501100002858, China Postdoctoral Science Foundation;
                Award ID: 2020T130109ZX
                Award ID: 2020T130609
                Funded by: National Key Research and Development Program of China;
                Award ID: 2018YFC2000500
                Funded by: COVID-19 Prevention and Control Program of International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University;
                Award ID: 2020-COVID-19-01
                Funded by: National S&T Major Project of China;
                Award ID: 2018ZX10301201
                Funded by: http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 82070643
                Award ID: U1904164
                Award ID: U2004121
                Funded by: Henan Province Science and Technology Project;
                Award ID: 202102310055
                Funded by: the opening foundation of the State Key Laboratory for Diagnosis and Treatment of Infectious Diseases and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases,The First Affiliated Hospital, College of Medicine, Zhejiang University;
                Award ID: SKLID2019KF03
                Categories
                Gut Microbiota
                1506
                2474
                2312
                Original research
                Custom metadata
                unlocked
                free

                Gastroenterology & Hepatology
                intestinal microbiology,lipid metabolism,covid-19
                Gastroenterology & Hepatology
                intestinal microbiology, lipid metabolism, covid-19

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