Detection of SARS Coronavirus in Patients with Severe Acute Respiratory Syndrome by Conventional and Real-Time Quantitative Reverse Transcription-PCR Assays

Poon, Leo L. M.; Chan, Kwok Hung; Wong, Otto; Cheung, Timothy; Ng, Iris H. Y.; Zheng, Bo‐Jian; Seto, W. H.; Yuen, Kwok‐Yung; Guan, Yi; Peiris, Malik

doi:10.1373/clinchem.2003.023663

Cited by 121 publications

(125 citation statements)

References 17 publications

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“…Only 1 in 3 piglets sheds detectable porcine respiratory CoV levels by 5 d following first clinical signs (Costantini et al 2004). Similarly, shedding of the human SARS virus peaks 7 to 10 d after onset of clinical signs (Poon et al 2004). It is possible that 4 HSCoV negative harbor seals had cleared the initial HSCoV infection or that the virus load in those lung samples had tapered beyond the detection limit of both our degenerate and specific PCR assay.…”

Section: Discussionmentioning

confidence: 88%

Detection of a respiratory coronavirus from tissues archived during a pneumonia epizootic in free-ranging Pacific harbor seals Phoca vitulina richardsii

Nollens¹,

Wellehan²,

Archer³

et al. 2010

Dis. Aquat. Org.

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In June 2000, 21 adult harbor seals Phoca vitulina richardsii were found dead along a localized section of the central California coast. Necropsy of 5 fresh carcasses revealed pulmonary congestion, consolidation, and hemorrhage. Histopathological changes in lungs from 2 of these seals included a necrotizing lymphocytic and histocytic lobar pneumonia with intra-lesional bacteria. A coronavirus (CoV) was detected in archived tissues from 1 of the 5 seals via a degenerate PCR for nidoviral RNA-dependent RNA polymerase (RdRp), and was subsequently confirmed via specific PCR. Based on the partial RdRp sequence, the CoV was identified as a novel, divergent member of the CoV group 1a. The virus is tentatively named harbor seal coronavirus (HSCoV). The clinical significance of HSCoV and its involvement in the etiology of the epizootic pneumonia and deaths of the harbor seals is uncertain. KEY WORDS: Harbor seal · Phoca vitulina richardsii · Coronavirus · Pneumonia Resale or republication not permitted without written consent of the publisherDis Aquat Org 90: [113][114][115][116][117][118][119][120] 2010 crochiropteran bat hosts; and Group 2d, from megachiropteran bat hosts (Woo et al. 2007). It has been proposed that Group 3 be divided as follows: Groups 3a, from avian hosts; Group 3b, which contains a coronavirus from a beluga whale Delphinapterus leucas, the only coronavirus identified in a marine mammal to date; and Group 3c, which contains viruses from diverse avian hosts as well as one from an Asian leopard cat Prionailurus bengalensis that may represent recent host switching (Woo et al. 2009).In late May and early June 2000, 21 adult harbor seals Phoca vitulina richardsii were found dead along a localized 10 mile (16 km) section of the central California coast at Point Reyes National Seashore. Upon postmortem examination of 5 relatively fresh carcasses, all seals had grossly abnormal lungs, and histological examination of 3 cases revealed severe pneumonia. A similar event with similar gross necropsy findings had occurred in 1997, when approximately 90 seals were found dead in the same area, but no conclusive etiology for the event was identified. In both events, a viral infection with secondary bacterial pneumonia was suspected. CoVs are recognized causes of enteric and respiratory infections that are often fatal in young animals. Here we report on the identification of a novel respiratory coronavirus from one of these harbor seals. This novel coronavirus of wildlife is a novel member of Group 1a and is distinct from, but most closely related to the CoVs of ferrets, cats, dogs, and swine. MATERIALS AND METHODS Animals and samples.On 26 May 2000, a harbor seal (HS D62) was found dead at McClure's Beach, Point Reyes National Seashore, in central California, and a field necropsy was conducted. Tonsil, stomach, cerebellum, brain stem, and lymph node were collected in 10% formalin for histological analysis. An additional 4 adult harbor seals were found dead on 6 June 2000. Two harbor seals, designated as H...

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Section: Discussionmentioning

confidence: 88%

Detection of a respiratory coronavirus from tissues archived during a pneumonia epizootic in free-ranging Pacific harbor seals Phoca vitulina richardsii

Nollens¹,

Wellehan²,

Archer³

et al. 2010

Dis. Aquat. Org.

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“…Without a gold diagnostic standard, it had been impossible to determine the usefulness of RT-PCR and serological testing for SARS-CoV. Nasopharyngeal aspirate and stool samples could be used for early diagnosis of SARS by detection of genomic RNA using real-time quantitative RT-PCR assay for SARS CoV, although the sensitivity is at most 80% [9]. RT-PCR systems to detect blood or serum SARS-CoV RNA provide a sensitivity of 50-87% during the first week of illness, but drop to 25% at day 14 after fever onset [10].…”

Section: Discussionmentioning

confidence: 99%

“…The use of RT-PCR, despite the initial optimism, does not permit complete confidence at early stages of the disease [5,[9][10], and takes 2-3 days for most microbiology laboratories to complete. Presence of anti-SARS-CoV IgG usually occurs earliest on day 10 of the illness, and serological detection is, therefore, not useful in the initial stages of SARS [5,11].…”

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confidence: 99%

A prediction rule for clinical diagnosis of severe acute respiratory syndrome

Ho¹,

Chau²,

Yip³

et al. 2005

Eur Respir J

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A prospective study was undertaken to identify clinical, radiographical, haematological and biochemical profiles of severe acute respiratory syndrome (SARS) patients.A prediction rule, which demarcates low from high risk patients for SARS in an outbreak situation was developed. A total of 295 patients with unexplained respiratory illnesses, admitted to Queen Mary Hospital, Hong Kong SAR, China, in March to July 2003, were evaluated for clinical, radiological, haematological and alanine transaminase (ALT) data daily for 3 days after hospitalisation.In total, 44 cases were subsequently confirmed to have SARS by RT-PCR (68.2%) and serology (100%). The scoring system of attributing 11, 10, 3, 3 and 3 points to the presence of independent risk factors, namely: epidemiological link, radiographical deterioration, myalgia, lymphopenia and elevated ALT respectively, generated high and low-risk (total score 11-30 and 0-10, respectively) groups for SARS. The sensitivity and specificity of this prediction rule in positively identifying a SARS patient were 97.7 and 81.3%, respectively. The positive and negative predictive values were 47.8 and 99.5%, respectively.The prediction rule appears to be helpful in assessing suspected patients with severe acute respiratory syndrome at the bedside, and should be further validated in other severe acute respiratory syndrome cohorts.

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“…The evaluated specimen types include nasopharyngeal aspirate (NPA) or swab, throat swab, faeces and urine [36][37][38][39][40][41][42][43]. In the upper respiratory tract, the viral load is low in the 7 days and peaks at around day 10 of illness [22,36,40]. Thus, diagnostic tests for SARS should be compared with reference to the timing of collection.…”

Section: Virological Diagnosis Of Severe Acute Respiratory Syndromementioning

confidence: 99%

“…Clinical experience has only been available on a few agents used in the 2003 outbreak, namely, ribavirin, Kaletra (ritonavir/lopinavir), interferons (IFN), corticosteroids, convalescence serum, and immunoglobulin (Ig). RT-PCR (nested) T/S (n=46) from suspected SARS Timing: NS; sensitivity: 37% [40] Real time RT-PCR NPA (n=170) Sensitivity: 26% (day 1-3), 41% (day 4-6), 62% (day 7-10) [36] Real time RT-PCR NPA, respiratory swabs, sputum, stool, urine (SARS: n=200; non-SARS: n=671)…”

Section: Pharmacotherapy Of Severe Acute Respiratory Syndromementioning

confidence: 99%

Diagnosis and pharmacotherapy of severe acute respiratory syndrome: what have we learnt?

Tsang¹

2004

Eur Respir J

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In 2003, the onset of severe acute respiratory syndrome (SARS) caused worldwide chaos. Although SARS was eradicated by isolation towards the end of 2003, sporadic cases have been reported in Singapore, Taiwan and mainland China.In this review, SARS is discussed as a disease, as well as its diagnosis, management and pharmacotherapy.Respiratory physicians and healthcare professionals have to be aware of advances in the understanding of the diagnosis and management of severe acute respiratory syndrome. More research is required in order to prepare for if this respiratory infection recurs, but there are concerns that adequate pharmaceutical support may be lacking for the development of a vaccine.

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Detection of SARS Coronavirus in Patients with Severe Acute Respiratory Syndrome by Conventional and Real-Time Quantitative Reverse Transcription-PCR Assays

Cited by 121 publications

References 17 publications

Detection of a respiratory coronavirus from tissues archived during a pneumonia epizootic in free-ranging Pacific harbor seals Phoca vitulina richardsii

Detection of a respiratory coronavirus from tissues archived during a pneumonia epizootic in free-ranging Pacific harbor seals Phoca vitulina richardsii

A prediction rule for clinical diagnosis of severe acute respiratory syndrome

Diagnosis and pharmacotherapy of severe acute respiratory syndrome: what have we learnt?

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