Diagnostic performance and clinical feasibility of a novel one-step RT-qPCR assay for simultaneous detection of multiple severe acute respiratory syndrome coronaviruses.

Coronavirus disease 2019 (COVID-19) is an acute respiratory infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Other coronaviruses (CoVs) can also infect humans, although the majority cause only mild respiratory symptoms. Because early diagnosis of SARS-CoV-2 is critical for preventing further transmission events and improving clinical outcomes, it is important to be able to distinguish SARS-CoV-2 from other SARS-related CoVs in respiratory samples. Therefore, we developed and evaluated a novel reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay targeting the genes encoding the spike (S) and membrane (M) proteins to enable the rapid identification of SARS-CoV-2, including several new circulating variants and other emerging SARS-like CoVs. By analysis of in vitro-transcribed mRNA, we established multiplex RT-qPCR assays capable of detecting 5 × 10° copies/reaction. Using RNA extracted from cell culture supernatants, our multiple simultaneous SARS-CoV-2 assays had a limit of detection of 1 × 10° TCID50/mL and showed no cross-reaction with human CoVs or other respiratory viruses. We also validated our method using human clinical samples from patients with COVID-19 and healthy individuals, including nasal swab and sputum samples. This novel one-step multiplex RT-qPCR assay can be used to improve the laboratory diagnosis of human-pathogenic CoVs, including SARS-CoV-2, and may be useful for the identification of other SARS-like CoVs of zoonotic origin.

Seroprevalence of Antibodies to SARS-CoV-2 in Guangdong Province, China between March to June 2020.

Guangdong province, located in South China, is an important economic hub with a large domestic migrant population and was among the earliest areas to report COVID-19 cases outside of Wuhan. We conducted a cross-sectional, age-stratified serosurvey to determine the seroprevalence of antibodies against SARS-CoV-2 after the emergence of COVID-19 in Guangdong. We tested 14,629 residual serum samples that were submitted for clinical testing from 21 prefectures between March and June 2020 for SARS-CoV-2 antibodies using a magnetic particle based chemiluminescent enzyme immunoassay and validated the results using a pseudovirus neutralization assay. We found 21 samples positive for SARS-CoV-2 IgG, resulting in an estimated age- and sex-weighted seroprevalence of 0.15% (95% CI: 0.06-0.24%). The overall age-specific seroprevalence was 0.07% (95% CI: 0.01-0.24%) in persons up to 9 years old, 0.22% (95% CI: 0.03-0.79%) in persons aged 10-19, 0.16% (95% CI: 0.07-0.33%) in persons aged 20-39, 0.13% (95% CI: 0.03-0.33%) in persons aged 40-59 and 0.18% (95% CI: 0.07-0.40%) in persons ≥60 years old. Fourteen (67%) samples had pseudovirus neutralization titers to S-protein, suggesting most of the IgG-positive samples were true-positives. Seroprevalence of antibodies to SARS-CoV-2 was low, indicating that there were no hidden epidemics during this period. Vaccination is urgently needed to increase population immunity to SARS-CoV-2.

The public health response to the COVID-19 outbreak in mainland China: a narrative review.

The COVID-19 pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused nations to adopt unprecedented control measures in order to curb its spread. As the first nation to respond, China's aggressive control measures appeared to have been effective in suppressing the first wave and keeping new cases under control. Here, we provide the historical context and details of China's public health response to COVID-19. We highlight the lessons and impact of the 2002-2003 SARS outbreak, which demonstrated the importance of transparency, surveillance and testing laboratories during an outbreak. We provide an overview of China's response strategy that was based on the principles of early detection, isolation, management and treatment and involved not only the large-scale coordination of multiple governmental bodies but also grass-root community participation throughout the country. These community-based organizations conducted active surveillance for febrile cases and provided support for those in quarantine and communities in lockdown. Importantly, these broader measures were supported by digital technology, including the extensive use of internet-based platforms and mobile applications (APPs). While there have been no significant increases in case numbers since April, there is still much concern over a second wave, considering the resumption of work and school, the lifting of travel restrictions and the outbreaks occurring globally. Control measures has since been implemented by provincial authorities, which includes continued surveillance and rapid testing. Although China's strict control measures may not suit every nation, the principles of early detection and isolation continue to hold true and have been a cornerstone of the initial and ongoing response to the COVID-19.

Human post-infection serological response to the spike and nucleocapsid proteins of SARS-CoV-2.

To inform seroepidemiological studies, we characterized the IgG- responses in COVID-19 patients against the two major SARS-CoV-2 viral proteins, spike (S) and nucleocapsid (N). We tested 70 COVID-19 sera collected up to 85 days post-symptom onset and 230 non-COVID-19 sera, including 27 SARS sera from 2003. Although the average SARS-CoV-2 S and N-IgG titers were comparable, N-responses were more variable among individuals. S- and N-assay specificity tested with non-COVID-19 sera were comparable at 97.5% and 97.0%, respectively. Therefore, S will make a better target due to its lower cross-reactive potential and its' more consistent frequency of detection compared to N.

SARS-CoV-2 environmental contamination associated with persistently infected COVID-19 patients.

BACKGROUND: Severe COVID-19 patients typically test positive for SARS-CoV-2 RNA for extended periods of time, even after recovery from severe disease. Due to the timeframe involved, these patients may have developed humoral immunity to SARS-CoV-2 while still testing positive for viral RNA in swabs. Data are lacking on exposure risks in these situations. Here, we studied SARS-CoV-2 environmental contamination in an ICU and an isolation ward caring for such COVID-19 patients. METHODS: We collected air and surface samples in a hospital caring for critical and severe COVID-19 cases from common areas and areas proximal to patients. RESULTS: Of the 218 ICU samples, an air sample contained SARS-CoV-2 RNA. Of the 182 isolation ward samples, nine contained SARS-CoV-2 RNA. These were collected from a facemask, the floor, mobile phones, and the air in the patient room and bathroom. Serum antibodies against SARS-CoV-2 were detected in these patients at the beginning of the study. CONCLUSIONS: While there is a perception of increased risk in the ICU, our study demonstrates that isolation wards may pose greater risks to healthcare workers and exposure risks remain with clinically improved patients, weeks after their initial diagnoses. As these patients had serum antibodies, further studies may be warranted to study the utility of serum antibodies as a surrogate of viral clearance in allowing people to return to work. We recommend continued vigilance even with patients who appear to have recovered from COVID-19.