Saliva as a diagnostic specimen for testing respiratory virus by a point-of-care molecular assay: a diagnostic validity study

To, Kelvin Kai-Wang; Yip, Cyril C. Y.; Lai, CL; Wong, Carlos King Ho; Ho, Deborah Tip Yin; Peng, Ping; Ng, Alexander; Leung, Kit Hang; Poon, Rosana W.S.; Chan, K.-H.; Cheng, Vincent Chi-Chung; Hung, Ivan Fan-Ngai; Yuen, Kwok‐Yung

doi:10.1016/j.cmi.2018.06.009

Cited by 180 publications

(178 citation statements)

References 25 publications

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“…If patients were intubated, we obtained endotracheal aspirate instead of posterior oropharynx saliva. 6,[9][10][11] Our initial experience showed that such saliva samples are promising in viral load monitoring in patients with COVID-19. 6 We also retrieved serum remnant from blood samples taken for routine bio chemical testing, and refrigerated these samples at -20°C until antibody testing could be done.…”

Section: Methodsmentioning

confidence: 99%

“…Findings of previous studies have shown high concordance between saliva and nasopharyngeal aspirate as specimens for laboratory diagnosis of respi ratory viruses. 9 We have reported use of posterior oro pharyngeal (deep throat) saliva for diagnosis and viral load monitoring in a cohort of 12 patients with COVID-19. 6 Here, we report use of self-collected posterior oro pharyn geal saliva samples from patients with COVID-19, which avoids close contact between healthcare workers and patients, for viral load monitoring.…”

Section: Introductionmentioning

confidence: 99%

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Temporal profiles of viral load in posterior oropharyngeal saliva samples and serum antibody responses during infection by SARS-CoV-2: an observational cohort study

Tsang²,

Leung³

et al. 2020

The Lancet Infectious Diseases

3,070

197

3,366

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Background Coronavirus disease 2019 (COVID-19) causes severe community and nosocomial outbreaks. Comprehensive data for serial respiratory viral load and serum antibody responses from patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are not yet available. Nasopharyngeal and throat swabs are usually obtained for serial viral load monitoring of respiratory infections but gathering these specimens can cause discomfort for patients and put health-care workers at risk. We aimed to ascertain the serial respiratory viral load of SARS-CoV-2 in posterior oropharyngeal (deep throat) saliva samples from patients with COVID-19, and serum antibody responses.Methods We did a cohort study at two hospitals in Hong Kong. We included patients with laboratory-confirmed COVID-19. We obtained samples of blood, urine, posterior oropharyngeal saliva, and rectal swabs. Serial viral load was ascertained by reverse transcriptase quantitative PCR (RT-qPCR). Antibody levels against the SARS-CoV-2 internal nucleoprotein (NP) and surface spike protein receptor binding domain (RBD) were measured using EIA. Whole-genome sequencing was done to identify possible mutations arising during infection.Findings Between Jan 22, 2020, and Feb 12, 2020, 30 patients were screened for inclusion, of whom 23 were included (median age 62 years [range 37-75]). The median viral load in posterior oropharyngeal saliva or other respiratory specimens at presentation was 5·2 log 10 copies per mL (IQR 4·1-7·0). Salivary viral load was highest during the first week after symptom onset and subsequently declined with time (slope -0·15, 95% CI -0·19 to -0·11; R²=0·71). In one patient, viral RNA was detected 25 days after symptom onset. Older age was correlated with higher viral load (Spearman's ρ=0·48, 95% CI 0·074-0·75; p=0·020). For 16 patients with serum samples available 14 days or longer after symptom onset, rates of seropositivity were 94% for anti-NP IgG (n=15), 88% for anti-NP IgM (n=14), 100% for anti-RBD IgG (n=16), and 94% for anti-RBD IgM (n=15). Anti-SARS-CoV-2-NP or anti-SARS-CoV-2-RBD IgG levels correlated with virus neutralisation titre (R²>0·9). No genome mutations were detected on serial samples.Interpretation Posterior oropharyngeal saliva samples are a non-invasive specimen more acceptable to patients and health-care workers. Unlike severe acute respiratory syndrome, patients with COVID-19 had the highest viral load near presentation, which could account for the fast-spreading nature of this epidemic. This finding emphasises the importance of stringent infection control and early use of potent antiviral agents, alone or in combination, for highrisk individuals. Serological assay can complement RT-qPCR for diagnosis.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Temporal profiles of viral load in posterior oropharyngeal saliva samples and serum antibody responses during infection by SARS-CoV-2: an observational cohort study

Tsang²,

Leung³

et al. 2020

The Lancet Infectious Diseases

3,070

197

3,366

View full text Add to dashboard Cite

show abstract

“…Respiratory samples of the patients were tested for influenza A and B viruses and respiratory syncytial virus using the Xpert Xpress Flu/RSV assay (GeneXpert System, Cepheid, Sunnyvale, CA, USA) according to the manufacturer's instructions. 6 To detect the presence of 18 respiratory virus targets and four bacteria (including adenovirus, coronaviruses [HCoV-229E, HCoV-Nl63, HCoV-Oc43, HCoV-HKU1, and MERS-CoV], human metapneumovirus, respiratory syncytial virus, human rhinovirus or entero virus, influenza A viruses [H1, H1-2009 and H3], influenza B virus, parainfluenza viruses [types 1 -4], Bordetella pertussis, Bordetella parapertussis, Chlamydophila pneumoniae, and Mycoplasma pneumoniae), samples were tested using BioFire FilmArray Respiratory Panel 2 plus (bioMérieux, Marcy l'Etoile, France) according to the manufacturer's instructions. 7 The two faecal samples were taken from the patients who had diarrhoea as part of their symptoms, and the samples were tested by BioFire FilmArray Gastrointestinal panel (bioMérieux) for 22 diarrhoeal pathogens.…”

Section: Respiratory and Diarrhoeal Pathogen Detectionmentioning

confidence: 99%

A familial cluster of pneumonia associated with the 2019 novel coronavirus indicating person-to-person transmission: a study of a family cluster

et al. 2020

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Background An ongoing outbreak of pneumonia associated with a novel coronavirus was reported in Wuhan city, Hubei province, China. Affected patients were geographically linked with a local wet market as a potential source. No data on person-to-person or nosocomial transmission have been published to date. MethodsIn this study, we report the epidemiological, clinical, laboratory, radiological, and microbiological findings of five patients in a family cluster who presented with unexplained pneumonia after returning to Shenzhen, Guangdong province, China, after a visit to Wuhan, and an additional family member who did not travel to Wuhan. Phylogenetic analysis of genetic sequences from these patients were done.Findings From Jan 10, 2020, we enrolled a family of six patients who travelled to Wuhan from Shenzhen between Dec 29, 2019 and Jan 4, 2020. Of six family members who travelled to Wuhan, five were identified as infected with the novel coronavirus. Additionally, one family member, who did not travel to Wuhan, became infected with the virus after several days of contact with four of the family members. None of the family members had contacts with Wuhan markets or animals, although two had visited a Wuhan hospital. Five family members (aged 36-66 years) presented with fever, upper or lower respiratory tract symptoms, or diarrhoea, or a combination of these 3-6 days after exposure. They presented to our hospital (The University of Hong Kong-Shenzhen Hospital, Shenzhen) 6-10 days after symptom onset. They and one asymptomatic child (aged 10 years) had radiological ground-glass lung opacities. Older patients (aged >60 years) had more systemic symptoms, extensive radiological ground-glass lung changes, lymphopenia, thrombocytopenia, and increased C-reactive protein and lactate dehydrogenase levels. The nasopharyngeal or throat swabs of these six patients were negative for known respiratory microbes by point-of-care multiplex RT-PCR, but five patients (four adults and the child) were RT-PCR positive for genes encoding the internal RNA-dependent RNA polymerase and surface Spike protein of this novel coronavirus, which were confirmed by Sanger sequencing. Phylogenetic analysis of these five patients' RT-PCR amplicons and two full genomes by nextgeneration sequencing showed that this is a novel coronavirus, which is closest to the bat severe acute respiatory syndrome (SARS)-related coronaviruses found in Chinese horseshoe bats.Interpretation Our findings are consistent with person-to-person transmission of this novel coronavirus in hospital and family settings, and the reports of infected travellers in other geographical regions.

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“…Since no invasive procedures are required, the collection of saliva can greatly minimize the chance of exposing healthcare workers to 2019-nCoV. We have previously demonstrated that saliva has a high concordance rate of greater than 90% with nasopharyngeal specimens in the detection of respiratory viruses, including coronaviruses [5,6]. In some patients, coronavirus was detected only in saliva but not in nasopharyngeal aspirate [5].…”

mentioning

confidence: 99%

Consistent Detection of 2019 Novel Coronavirus in Saliva

Tsang²,

Yip

et al. 2020

Clinical Infectious Diseases

1,691

1,683

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Saliva as a diagnostic specimen for testing respiratory virus by a point-of-care molecular assay: a diagnostic validity study

Cited by 180 publications

References 25 publications

Temporal profiles of viral load in posterior oropharyngeal saliva samples and serum antibody responses during infection by SARS-CoV-2: an observational cohort study

Temporal profiles of viral load in posterior oropharyngeal saliva samples and serum antibody responses during infection by SARS-CoV-2: an observational cohort study

A familial cluster of pneumonia associated with the 2019 novel coronavirus indicating person-to-person transmission: a study of a family cluster

Consistent Detection of 2019 Novel Coronavirus in Saliva

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