ABSTRACT
Timely and accurate detection of SARS-CoV-2 variants of concern (VOCs) is urgently needed for pandemic surveillance and control. Great efforts have been made from a mass of scientists in increasing the detection sensitivity and operability, and reducing the turn-around time and cost. Here, we report a nucleic acid testing-based method aiming to detect and discriminate SARS-CoV-2 mutations by combining RT-RPA and CRISPR-Cas12a detecting assays (RRCd). With a detection limit of 10 copies RNA/reaction, RRCd was validated in 194 clinical samples, showing 89% positive predictive agreement and 100% negative predictive agreement, respectively. Critically, using specific crRNAs, representatives of single nucleotide polymorphisms and small deletions in SARS-CoV-2 VOCs including N501Y, T478K and ΔH69-V70 were discriminated by RRCd, demonstrating 100% specificity in clinical samples with C t < 33. The method completes within 65 min and could offer visible results without using any electrical devices, which probably facilitate point-of-care testing of SARS-CoV-2 variants and other epidemic viruses.
ABSTRACT
BACKGROUND: Global mobility of the population has accelerated spread of the Human Norovirus (HuNoV), with long-distance travel in enclosed spaces increasing the opportunity for viral outbreaks. However, surveillance of HuNoV transmission is still lacking, especially in cross-border transportation. METHOD: From 533 self-reported patients, 83 swab samples (15.6%) tested positive for HuNoV by RT-qPCR. Positive samples were sequenced using next-generation sequencing (NGS). Epidemiological investigation and whole genome analysis were then conducted. RESULTS: Most cases occurred in February and March, with large outbreaks involving more than 34 people. A total of 74 HuNoV sequences that could be genotyped were obtained, with near-complete genomes (>7 kb) accounting for most sequences (57/74). A total of 19 different genotypes of viral whole genome sequences were included. The first whole genome sequence of GII.9[P7] was obtained. Rarely reported genotypes including GI.3[P10], GI.3[P13], GII.7[P7], GII.8[P8], and GIX.1[GII.P15] were sequenced and assembled successfully. Four possible sources of virus outbreaks in China were traced. Beyond HuNoV, whole genome sequences of food-borne viruses including Salivirus, Kobuvirus, and Enterovirus were obtained in further assembly. CONCLUSIONS: Surveillance of the etiology and epidemiology of HuNoV global spread through travelers will improve pre-travel health advice, empirical treatment, and estimates of vaccine-preventable diseases.
