The SARS-CoV-2 Omicron variant sparked the largest wave of infections worldwide. Mainland China eased its strict COVID-19 measures in late 2022 and experienced two nationwide Omicron waves in 2023. Here, we investigated lineage distribution and virus evolution in Guangdong, China, 2022-2023 by comparing 5813 local viral genomes with the datasets from other regions of China and worldwide. Additionally, we conducted three large-scale serological surveys involving 1696 participants to measure their immune response to the BA.5 and XBB.1.9 before and after the corresponding waves. Our findings revealed the Omicron variants, mainly the BA.5.2.48 lineage, causing infections in over 90% of individuals across different age groups within a month. This rapid spread led to the establishment of widespread immunity, limiting the virus’s ability to further adaptive mutation and dissemination. While similar immune responses to BA.5 were observed across all age groups after the initial wave, children aged 3 to 11 developed a stronger cross immune response to the XBB.1.9 strain, possibly explaining their lower infection rates in the following XBB.1 wave. Reinfection with Omicron XBB.1 variant triggered a more potent neutralizing immune response among older adults. These findings highlight the impact of age-specific immune responses on viral spread in potential future waves.

Background: In 2024, mainland China witnessed a significant upsurge in Hand, Foot, and Mouth Disease (HFMD) cases. Coxsackievirus A16 (CVA16) is one of the primary causative agents of HFMD. Long-term monitoring of theCVA16 infection rate and genotype changes is crucial for the prevention and control of HFMD. Methods: A total of 40,673 clinical specimens were collected from suspected HFMD cases in Guangdong province from 2018 to 2024, including rectal swabs (n = 27,954), throat swabs (n = 6791), stool (n = 5923), cerebrospinal fluid (n = 3), and herpes fluid (n = 2). A total of 24,410 samples were detected as EV-positive and further typed by RT-PCR. A total of 872 CVA16-positive samples were isolated and further sequenced to obtain the full-length VP1 sequence. Phylogenetic analysis was performed based on viral protein 1 gene (VP1). Results: In the first 25 weeks of 2024, reported cases of HFMD were 1.36 times higher than the mean rates of 2023. In 2024, CVA16 predominated at 75.42%, contrasting with the past etiological pattern in which the CVA6 was predominant with the detection rate ranging from 32.85 to 77.61% from 2019 to 2023. Phylogenetic analysis based on the VP1 gene revealed that the B1a and B1b subtypes co-circulated in Guangdong from 2018 to 2022. The B1c outbreak clade, detected in Guangdong in 2023, constituted 68.24% of the 148 strains of CVA16 collected in 2024, suggesting a subtype shift in the CVA16 virus. There were three specific amino acid variations (P3S, I235V, and T240A) in the VP1 sequence of B1c. Conclusions: The new emergence of the CVA16 B1c outbreak clade in Guangdong during 2023–2024 highlights the necessity for the enhanced surveillance of the virus evolution epidemiological dynamic in this region. Furthermore, it is imperative to closely monitor the etiological pattern changes in Hand, Foot, and Mouth Disease (HFMD) in other regions as well. Such vigilance will be instrumental in guiding future vaccination strategies for HFMD.

Following the normalization of the COVID-19 pandemic, the focus of wastewater-based epidemiology (WBE) must be broadened from SARS-CoV-2 to encompass surveillance of other major infectious diseases, particularly for pathogens where conventional clinical monitoring systems exhibit inherent surveillance gaps. In this study, we conducted a continuous two-year WBE study (January 2023 to December 2024) across three high-population-density cities in Guangdong, China to establish epidemiological baselines for enteric diarrheal viruses. We analyzed monthly raw wastewater samples from major treatment plants using advanced molecular methods, including digital PCR (ddPCR) for viral load quantification and targeted high-throughput sequencing (tNGS) for genotypic analysis. Our findings revealed diverse circulation patterns among the monitored enteric viruses. Astrovirus (AstV) had the highest detection rate (100%), reflecting its broad endemic distribution, while Norovirus genogroup II (NoV GII) exhibited relatively high viral loads (median 4 × 10⁴ copies/mL) and presented explosive seasonal peaks (significant upward trend in spring.), highlighting its epidemic potential. Furthermore, distinct spatiotemporal patterns were observed, with Sapovirus showing a significant summer peak in Foshan city, contrasting with the winter/spring peaks in the other cities. The tNGS results demonstrated similar sensitivity to RT-PCR in virus detection, and sequencing analyses uncovered the co-circulation and periodic shifts in dominant viral genotypes, such as the emergence of multiple NoV and AstV lineages. This longitudinal WBE surveillance successfully established critical baseline data and demonstrated significant regional heterogeneity in viral circulation, providing essential, complementary data to inform public health strategies for preventing diarrheal outbreaks in urban settings.