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An Overview of the Temporal Shedding of SARS-CoV-2 RNA in Clinical Specimens.

ABSTRACT: Coronavirus disease 2019 quickly spread in China and has, since March 2020 become a pandemic, causing hundreds of thousands of deaths worldwide. The causative agent was promptly isolated and named SARS-CoV-2. Scientific efforts are related to identifying the best clinical management of these patients, but also in understanding their infectivity in order to limit the spread of the virus. Aimed at identifying viral RNA in the various compartments of the organism of sick subjects, diagnostic tests are carried out. However, the accuracy of such tests varies depending on the type of specimen used and the time of illness at which they are performed. This review of the literature aims to summarize the preliminary findings reported in studies on Covid-19 testing. The results highlight how the pharyngeal swab is highly sensitive in the first phase of the disease, while in the advanced stages, other specimens should be considered, such as sputum, or even stool to detect SARS-CoV-2. It highlights that most patients already reach the peak of the viral load in the upper airways within the first days of displaying symptoms, which thereafter tend to decrease. This suggests that many patients may already be infectious before symptoms start to appear.

SUBMITTER: Zhurakivska K

PROVIDER: S-EPMC7468374 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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An Overview of the Temporal Shedding of SARS-CoV-2 RNA in Clinical Specimens.

Zhurakivska Khrystyna K Troiano Giuseppe G Pannone Giuseppe G Caponio Vito Carlo Alberto VCA Lo Muzio Lorenzo L

Frontiers in public health 20200820

Coronavirus disease 2019 quickly spread in China and has, since March 2020 become a pandemic, causing hundreds of thousands of deaths worldwide. The causative agent was promptly isolated and named SARS-CoV-2. Scientific efforts are related to identifying the best clinical management of these patients, but also in understanding their infectivity in order to limit the spread of the virus. Aimed at identifying viral RNA in the various compartments of the organism of sick subjects, diagnostic tests ...[more]

PMID: 32974267

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Project description:SARS-CoV-2 transcribes a set of subgenomic RNAs (sgRNAs) essential for the translation of structural and accessory proteins to sustain its life cycle. We applied RNA-seq on 375 respiratory samples from individual COVID-19 patients and revealed that the majority of the sgRNAs were canonical transcripts with N being the most abundant (36.2%), followed by S (11.6%), open reading frame 7a (ORF7a; 10.3%), M (8.4%), ORF3a (7.9%), ORF8 (6.0%), E (4.6%), ORF6 (2.5%), and ORF7b (0.3%); but ORF10 was not detected. The profile of most sgRNAs, except N, showed an independent association with viral load, time of specimen collection after onset, age of the patient, and S-614D/G variant with ORF7b and then ORF6 being the most sensitive to changes in these characteristics. Monitoring of 124 serial samples from 10 patients using sgRNA-specific real-time RT-PCR revealed a potential of adopting sgRNA as a marker of viral activity. Respiratory samples harboring a full set of canonical sgRNAs were mainly collected early within 1 to 2 weeks from onset, and most of the stool samples (90%) were negative for sgRNAs despite testing positive by diagnostic PCR targeting genomic RNA. ORF7b was the first to become undetectable and again being the most sensitive surrogate marker for a full set of canonical sgRNAs in clinical samples. The potential of using sgRNA to monitor viral activity and progression of SARS-CoV-2 infection, and hence as one of the objective indicators to triage patients for isolation and treatment should be considered. IMPORTANCE Attempts to use subgenomic RNAs (sgRNAs) of SARS-CoV-2 to identify active infection of COVID-19 have produced diverse results. In this work, we applied next-generation sequencing and RT-PCR to profile the full spectrum of SARS-CoV-2 sgRNAs in a large cohort of respiratory and stool samples collected throughout infection. Numerous known and novel discontinuous transcription events potentially encoding full-length, deleted and frameshift proteins were observed. In particular, the expression profile of canonical sgRNAs was associated with genomic RNA level and clinical characteristics. Our study found sgRNAs as potential biomarkers for monitoring infectivity and progression of SARS-CoV-2 infection, which provides an alternative target for the management and treatment of COVID-19 patients.

Project description:SARS-CoV-2 shedding in human stool has been suggested as a probable route for faeco-oral transmission of the virus due to the availing evidence on the infectivity and pathogenicity of similar highly infectious respiratory viruses. Determining association of SARS-CoV-2 shedding in stools and presenting clinical status might be useful for prediction of the viral transmission spectrum and disease outcome. This study involved a descriptive cross-sectional survey of 91 consenting hospitalized, confirmed COVID-19 patients in Infectious Disease Isolation Centre, Oyo State, Nigeria. Socio-demographic characteristics and other ancillary data were collected from patient's hospital records with the aid of a structured investigator administered questionnaire. The laboratory detection of SARS-CoV-2 RNA in the stool of patients was performed using RT-PCR method. 27 (29.7%) of the 91 COVID-19 patients shed SARS-CoV-2 in their stools. The frequency of male (38.3%) patients shedding the virus in stools was higher than female (12.9%) patients (P = 0.012). Higher proportion of patients who had symptoms (41.2%) at admission shed the virus in their stool (P <0.007); particularly, fever (0.001), fatigue (0.003), headache (0.003), catarrh (0.001), and loss of smell (0.009). The frequency of viral shedding in stool was higher among patients with loss of taste (p = 0.028). Viral shedding in stool was significantly associated with low CT values (47.2%) and moderate CT value (21.4%) (P<0.05). Multivariate analysis showed that patients with moderate CT-value (OR = 0.28, 95% CI: 0.08-0.94, P = 0.039) and high CT-value (OR = 0.08, 95% CI: 0.01-0.80, P = 0.033) were less likely to shed the virus in stool. The gastrointestinal tract could be a route of SARS-CoV-2 transmission irrespective of the patients' clinical status. The low and moderate CT values of the nasopharyngeal swab is associated with shedding of the virus in patients' stools, although infectivity will depend on viral activity obtainable from further laboratory test analyses, such as viral culture.

Dataset Information

An Overview of the Temporal Shedding of SARS-CoV-2 RNA in Clinical Specimens.

Publications

An Overview of the Temporal Shedding of SARS-CoV-2 RNA in Clinical Specimens.

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