Validation and implementation of a direct RT-qPCR method for rapid screening of SARS-CoV-2 infection by using non-invasive saliva samples.

OBJECTIVE: To validate and implement an optimized screening method for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA combining use of self-collected raw saliva samples, single-step heat-treated virus inactivation and RNA extraction, and direct RT-qPCR. METHODS: This was a three-phase study conducted in Barcelona (Spain) during June to October, 2020. The three phases were (1) analytical validation against standard RT-qPCR in saliva samples; (2) diagnostic validation against standard RT-qPCR using paired saliva-nasopharyngeal samples obtained from asymptomatic teenagers and adults in a sports academy; and (3) pilot screening of asymptomatic health workers in a tertiary hospital. RESULTS: In phase 1, the detection yield of the new method was comparable to that of standard RT-qPCR. In phase 2, the diagnostic sensitivity and specificity values in 303 self-collected saliva samples were 95.7% (95% confidence interval 79.0-99.2%) and 100.0% (95% confidence interval 98.6-100.0%), respectively. In phase 3, only 17 (0.6%) of the saliva samples self-collected by 2709 participants without supervision were invalid. The rapid analytical workflow with the new method (up to 384 batched samples could be processed in less than 2 hours) yielded 24 (0.9%) positive results in the remaining 2692 saliva samples. Paired nasopharyngeal specimens were all positive by standard RT-qPCR. CONCLUSIONS: Direct RT-qPCR on self-collected raw saliva is a simple, rapid, and accurate method with potential to be scaled up for enhanced SARS-CoV-2 community-wide screening.

Performance Comparison of a Novel Rapid Stand-alone Molecular Test and a 2-Step Diagnostic Algorithm for Clostridioides difficile Detection in Children.

BACKGROUND: We aimed to evaluate diagnostic performance of the cobas® Liat® Cdiff test, a novel single-step automated polymerase chain reaction (PCR) assay for rapid diagnosis of toxigenic Clostridioides difficile infection (CDI) in stool samples from children with clinical symptoms of CDI. METHODS: Assessment of cobas Liat Cdiff diagnostic yield, time of analytical process and agreement of results with those of a 2-step diagnostic algorithm. The sequential algorithm combined an enzyme immunoassay (EIA) targeting antigen glutamate dehydrogenase (GDH), enterotoxin-A and cytotoxin-B, and a confirmatory PCR in EIA GDH-positive and toxin-negative samples. Fresh stool samples were collected prospectively from patients 2-18 years of age that were attended in Hospital Sant Joan de Deu (Barcelona, Spain) during December 2018-August 2019. RESULTS: A total of 122 specimens were collected from 91 children (mean age, 8 years; 69.7% male). cobas Liat Cdiff identified 24 (19.7%) positive samples. EIA yielded 97 (79.5%) GDH- and toxin-negative results, 11 (9.0%) GDH- and toxin-positive results, and 14 (11.5%) GDH-positive and toxin-negative results, of which 11 (9.0%) were positive for the toxin by the confirmatory PCR. Overall, GDH- and toxin-positive samples detected by the sequential algorithm were 22 (18.0%). Comparatively, the new test reduced time of the analytical process significantly (20 vs. 35.4 minutes, P < 0.001). CONCLUSION: Use of cobas Liat Cdiff showed similar detection yield compared with a 2-step diagnostic algorithm that combined an EIA and a confirmatory PCR while decreasing the time of the analytical process markedly in stool samples from children suspected of CDI.