Identification of a Novel Coronavirus in Patients with Severe Acute Respiratory Syndrome

TLDR

Severe acute respiratory syndrome (SARS) has recently been identified as a new clinical entity. The authors isolated a novel coronavirus from SARS patients using cell culture, sequenced a 300‑nt region via PCR‑based random amplification, and developed conventional and real‑time PCR assays for its detection. A novel coronavirus, distantly related to known coronaviruses, was isolated from SARS patients, detected in multiple specimen types—including sputum with up to 100 million RNA copies/mL, plasma during the acute phase, and feces during convalescence—while seroconversion occurred, indicating a likely etiologic role in SARS.

Abstract

BackgroundAbstractThe severe acute respiratory syndrome (SARS) has recently been identified as a new clinical entity. SARS is thought to be caused by an unknown infectious agent.MethodsClinical specimens from patients with SARS were searched for unknown viruses with the use of cell cultures and molecular techniques.ResultsA novel coronavirus was identified in patients with SARS. The virus was isolated in cell culture, and a sequence 300 nucleotides in length was obtained by a polymerase-chain-reaction (PCR)–based random-amplification procedure. Genetic characterization indicated that the virus is only distantly related to known coronaviruses (identical in 50 to 60 percent of the nucleotide sequence). On the basis of the obtained sequence, conventional and real-time PCR assays for specific and sensitive detection of the novel virus were established. Virus was detected in a variety of clinical specimens from patients with SARS but not in controls. High concentrations of viral RNA of up to 100 million molecules per milliliter were found in sputum. Viral RNA was also detected at extremely low concentrations in plasma during the acute phase and in feces during the late convalescent phase. Infected patients showed seroconversion on the Vero cells in which the virus was isolated.ConclusionsThe novel coronavirus might have a role in causing SARS.