Evaluation of nested polymerase chain methods for the detection of human coronaviruses 229E and OC43

Myint, S.; Johnston, Sebastian L.; Sanderson, G.; Simpson, Helen

doi:10.1006/mcpr.1994.1052

Cited by 86 publications

(85 citation statements)

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“…1C). We isolated RNA from the supernatant of Vero cells infected with the unidentified virus isolate for RT-PCR analyses using primer sets specific for the known human coronaviruses OC43 and 229E (6). Whereas the control viruses reacted positively with the respective virus-specific primers, the previously undescribed virus isolate failed to do so (data not shown).…”

Section: Resultsmentioning

confidence: 99%

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A previously undescribed coronavirus associated with respiratory disease in humans

Fouchier

Hartwig

Bestebroer

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

528

498

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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.

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Section: Resultsmentioning

confidence: 99%

“…RNA was isolated from infected Vero cell supernatants by using a High Pure RNA Isolation kit according to instructions from the manufacturer (Roche Diagnostics, Almere, The Netherlands). RT-PCR assays for HCoV-OC43, HCoV-229E were performed as described (6). Broad reacting primer sets specific for the Coronaviridae family were also used (7,8).…”

Section: Methodsmentioning

confidence: 99%

A previously undescribed coronavirus associated with respiratory disease in humans

Fouchier

Hartwig

Bestebroer

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

528

498

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“…Firstly, viral ribonucleic acid (RNA) was identified by reverse transcriptasepolymerase chain reaction (RT-PCR) in nasal lavage fluid, and throat and nose swabs taken on days 2 and 4 postinoculation. This method was derived from a nested RT-PCR described previously, which has been validated as being more sensitive than tissue culture [20]. The throat swab was taken from the posterior pharynx and tonsil bed, and the nose swab from the inferior turbinate.…”

Section: Confirmation Of Viral Infectionmentioning

confidence: 99%

“…The RT-PCR is highly specific with primers designed to target the N-gene of HCoV only [20]. False-positives from contamination during the RT-PCR assay are unlikely as none were detected in the duplicate reverse transcript negative control samples.…”

Section: Evidence Of Viral Infectionmentioning

confidence: 99%

“…False-positives from contamination during the RT-PCR assay are unlikely as none were detected in the duplicate reverse transcript negative control samples. RT-PCR is a highly sensitive and specific assay that for HCoV detection is superior to serology [20]. This is borne out in a Southampton study of 292 reported virus induced exacerbations of asthma suffered by 108 children aged 9 -11 yrs [14].…”

Section: Evidence Of Viral Infectionmentioning

confidence: 99%

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A model of viral wheeze in nonasthmatic adults: symptoms and physiology

McKean

Leech²,

Lambert³

et al. 2001

Eur Respir J

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Episodic wheezing associated with viral infections of the upper respiratory tract (URT) is a common problem in young children but also occurs in adults. It is hypothesized that an experimental infection with human coronavirus (HCoV), the second most prevalent common cold virus, would cause lower respiratory tract (LRT) changes in adults with a history of viral wheeze.Twenty-four viral wheezers (15 atopic) and 19 controls (seven atopic) were inoculated with HCoV 229E and monitored for the development of symptoms, changes in airway physiology and provocative concentration of methacholine causing a 20% fall in forced expiratory volume in one second (FEV1) (PC20). At baseline, viral wheezers were similar to controls in PC20 (mean¡SD log 2 PC20: 5.1¡1.9 and 5.8¡1.4 g?L -1 , respectively) but had a lower FEV1 than controls (mean¡SD 85.8¡11.4 and 95.6¡13.2% predicted, respectively pv0.05). Nineteen viral wheezers and 11 controls developed colds. Viral wheezers with colds reported significantly more URT symptoms than controls (median scores (interquartile range): 24 (10 -37) and 6 (4 -15), respectively p~0.014). Sixteen viral wheezers and no controls reported LRT symptoms (wheeze, chest tightness and shortness of breath). The viral wheezers with colds had small (3 -4%) reductions in FEV1 and peak expiratory flow on days with LRT symptoms (days 3 -6), but a progressive reduction in PC20 from baseline on days 2, 4 and 17 after inoculation (by 0.82, 1.35 and 1.82 doubling concentrations, respectively). The fall in PC20 affected both atopic and nonatopic subjects equally. There were no changes in FEV1 or PC20 in controls.An adult model of viral wheeze that is independent of atopy and therefore, of classical atopic asthma was established.

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Effect of Daily Vitamin E and Multivitamin-Mineral Supplementation on Acute Respiratory Tract Infections in Elderly Persons

Graat

Schouten²,

Kok³

2002

JAMA

218

193

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Evaluation of nested polymerase chain methods for the detection of human coronaviruses 229E and OC43

Cited by 86 publications

References 0 publications

A previously undescribed coronavirus associated with respiratory disease in humans

A previously undescribed coronavirus associated with respiratory disease in humans

A model of viral wheeze in nonasthmatic adults: symptoms and physiology

Effect of Daily Vitamin E and Multivitamin-Mineral Supplementation on Acute Respiratory Tract Infections in Elderly Persons

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