A previously undescribed coronavirus associated with respiratory disease in humans

Abstract

The etiology of acute respiratory tract illnesses is sometimes unclear due to limitations of diagnostic tests or the existence of as-yet-unidentified pathogens. Here we describe the identification and characterization of a not previously recognized coronavirus obtained from an 8-mo-old boy suffering from pneumonia. This coronavirus replicated efficiently in tertiary monkey kidney cells and Vero cells, in contrast to human coronaviruses (HCoV) 229E and OC43. The entire cDNA genome sequence of the previously undescribed coronavirus was determined, revealing that it is most closely related to porcine epidemic diarrhea virus and HCoV 229E. The maximum amino acid sequence identity between ORFs of the newly discovered coronavirus and related group 1 coronaviruses ranged from 43% to 67%. Real-time RT-PCR assays were designed to test for the prevalence of the previously undescribed coronavirus in humans. Using these tests, the virus was detected in four of 139 individuals (3%) who were suffering from respiratory illness with unknown etiology. All four patients suffered from fever, runny nose, and dry cough, and all four had underlying or additional morbidity. Our data will enable the development of diagnostic tests to study the prevalence and clinical impact of this virus in humans in more detail. Moreover, it will be important to discriminate this previously undescribed coronavirus from HCoV 229E and OC43 and the severe acute respiratory syndrome coronavirus.

Severe acute respiratory syndrome coronavirus-like virus in Chinese horseshoe bats

“…Primers (5Ј-CGCGGATCCGATGTCTGATAATG-GACCC-3Ј and 5Ј-CGGAATTCTTATGCCTGAGTAGAA-TCA-3Ј) were used to amplify the N gene of bat-SARS-CoV by RT-PCR. Western blot analysis, using 900 ng of purified bat-SARS-CoV (His) 6 -tagged N protein and sera at 1:1,000 dilution, was performed as described in ref. 22.…”

“…Prediction of signal peptides and cleavage sites was performed by using SIGNALP software, transmembrane domains by using TMPRED and TMHMM software, potential N-glycosylation sites by using SCANPROSITE software, and protein family analysis by using PFAM and INTERPROSCAN software (15)(16)(17)(18)(19)(20)(21). Cloning and purification of (His) 6 -tagged recombinant N protein of bat-SARS-CoV were performed as described in ref. 22.…”

“…22. Nunc immunoplates coated with 20 ng of purified (His) 6 -tagged recombinant bat-SARS-CoV N protein per well were used. Detection was performed by using 1:2,000 horseradish peroxidase-conjugated protein G and 3,3Ј,5,5Ј-tetramethylbenzidine, both from Zymed.…”

“…Their tendency for recombination and high mutation rates may allow them to adapt to new hosts and ecological niches. Among CoV that infect humans, including human CoV 229E (HCoV-229E), human CoV OC43 (HCoV-OC43), severe acute respiratory syndrome CoV (SARS-CoV), human CoV NL63 (HCoV-NL63), and CoV HKU1 (CoV-HKU1), SARS-CoV causes the most severe disease, with Ͼ700 fatalities reported since the SARS epidemic in 2003 (2)(3)(4)(5)(6)(7)(8).…”

Coronavirus Infections in Pediatric Outpatients with Febrile Respiratory Tract Infections in Hiroshima, Japan, over a 3-Year Period

“…Two of the HCoV strains (HCoV-OC43 and HCoV-229E) were detected predominantly in individuals with common colds (3)(4)(5)(6). Thereafter, three new HCoV strains were identified via molecular and tissue culture assays: severe acute respiratory syndrome coronavirus (SARS-CoV) (7), coronavirus strain noted in The Netherlands (HCoV-NL63) (8,9), and coronavirus strain noted in Hong Kong (HCoV-HKU1) (10). SARSCoV was associated with an outbreak of severe pneumonia that spread from Asia to other parts of the world, but since then has not caused any outbreaks.…”