RespiCoV: Simultaneous identification of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and 46 respiratory tract viruses and bacteria by amplicon-based Oxford-Nanopore MinION sequencing

PLoS One. 2022 Mar 9;17(3):e0264855. doi: 10.1371/journal.pone.0264855. eCollection 2022.

Abstract

Since December 2019 the world has been facing the outbreak of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Identification of infected patients and discrimination from other respiratory infections have so far been accomplished by using highly specific real-time PCRs. Here we present a rapid multiplex approach (RespiCoV), combining highly multiplexed PCRs and MinION sequencing suitable for the simultaneous screening for 41 viral and five bacterial agents related to respiratory tract infections, including the human coronaviruses NL63, HKU1, OC43, 229E, Middle East respiratory syndrome coronavirus, SARS-CoV, and SARS-CoV-2. RespiCoV was applied to 150 patient samples with suspected SARS-CoV-2 infection and compared with specific real-time PCR. Additionally, several respiratory tract pathogens were identified in samples tested positive or negative for SARS-CoV-2. Finally, RespiCoV was experimentally compared to the commercial RespiFinder 2SMART multiplex screening assay (PathoFinder, The Netherlands).

MeSH terms

  • Bacteria / genetics*
  • Bacteria / isolation & purification
  • COVID-19 / diagnosis*
  • COVID-19 / virology
  • Coronavirus / genetics
  • Coronavirus / isolation & purification
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / metabolism
  • Herpesvirus 1, Human / genetics
  • Herpesvirus 1, Human / isolation & purification
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Multiplex Polymerase Chain Reaction
  • Nanopores
  • Orthomyxoviridae / genetics
  • Orthomyxoviridae / isolation & purification
  • RNA Viruses / genetics*
  • RNA Viruses / isolation & purification
  • RNA, Viral / chemistry
  • RNA, Viral / metabolism
  • Respiratory Tract Infections / diagnosis*
  • Respiratory Tract Infections / microbiology
  • Respiratory Tract Infections / virology
  • SARS-CoV-2 / genetics*
  • SARS-CoV-2 / isolation & purification

Substances

  • DNA, Bacterial
  • RNA, Viral

Grants and funding

The author(s) received no specific funding for this work.