During the early 2025/26 influenza season, influenza A(H3N2) subclade K rapidly predominated in Beijing, China. Using a test-negative design, we estimated influenza vaccine effectiveness (VE) among influenza-like illness outpatients tested between weeks 40/2025 and 04/2026. Among 10,484 participants, sequencing of 316 randomly selected A(H3N2)-positive samples showed 84.8% were subclade K, and antigenic analysis of 65 viruses indicated antigenic divergence. Despite this, adjusted VE against laboratory-confirmed influenza was 23.5% (95% confidence interval: 11.7–33.7), indicating modest protection during this subclade K-dominated season.

OBJECTIVES:Schools are high-risk settings for influenza transmission. During the 2025-26 Northern Hemisphere season, influenza A(H3N2) subclade K (J.2.4.1) rapidly emerged and became predominant, but vaccine effectiveness (VE) in school-associated outbreaks remains unclear. METHODS:We conducted a field investigation of 93 school-associated febrile outbreaks in Beijing, China, from September to December 2025. Cases were febrile students with laboratory-confirmed influenza, and afebrile classmates served as controls. VE against laboratory-confirmed symptomatic infection was estimated using mixed-effects logistic regression, adjusting for age, sex, school type, and outbreak-level clustering. RESULTS:A total of 3797 students were included, of whom 788 were laboratory-confirmed febrile cases and 3009 were afebrile classmates. Primary schools accounted for 74.2% of outbreaks. Overall vaccination rate was 54.6%, with 50.6% among cases and 56.1% among controls. Unadjusted VE was 19.6% (95% CI: 5.9-31.3). After adjustment, VE against laboratory-confirmed influenza was 30.0% (95% CI: 17.3-40.7). Sensitivity analyses yielded consistent results. CONCLUSION:Influenza vaccination provided modest protection against symptomatic A(H3N2) infection in school-aged children, even under conditions of high-intensity exposure and antigenic drift. These findings underscore the value of vaccination in outbreak-prone school settings and highlight the importance of ongoing vaccine strain optimization and outbreak-based VE surveillance.

Human adenovirus B7 (HAdV-B7) has emerged as an increasingly important respiratory pathogen in recent years. Ten first grade students experienced symptoms of acute respiratory infection at a primary school in Beijing from November 13 to 21, 2024. We investigated the cause of this outbreak. In partnership with the local center for disease prevention and control, we collected demographic information (age and gender) of the affected students and obtained pharyngeal swab specimens from six students. We used multiplex real-time PCR to screen for 22 common respiratory viruses in these samples. To isolate HAdV, Hep-2 cells were inoculated with HAdV-positive pharyngeal swab samples. Isolates were sequenced by Sanger and whole-genome sequencing. In addition, four HAdV-B7 strains isolated from 2024 to 2025 influenza-like illness surveillance cases were also sequenced as a control to determine whether the same mutations existed. The strains were characterized molecularly using gene homology analysis, recombination analysis and phylogenetic reconstruction. We collected six pharyngeal swab samples from ten students based on consent. All samples were HAdV-positive as determined by multiplex real-time PCR. None of the students were co-infected with other viruses. We successfully isolated the HAdV strain from each student sample. Blast analysis and phylogenetic analysis of the hexon, penton, and fiber genes suggested that this outbreak was caused by HAdV-B7 (P7H7F7). A unique T335S mutation was identified within the RGD loop of the penton base in this outbreak. Whole-genome sequencing confirmed high sequence identity between our isolates and HAdV-B7 strains from multiple regions in China and abroad. In addition, all strains obtained in this outbreak had 57 nucleotides deletion in the E3 region. Interestingly, the same deletion mutants were also found in HAdV-B7 strains isolated from 2024 to 2025 influenza-like illness surveillance cases, indicating that this deletion mutation has been prevalent and may represent an established molecular signature of a circulating HAdV-B7 sublineage. We report the first detection of a human adenovirus type 7 deletion mutant associated with an outbreak of acute respiratory infections in Beijing in 2024. This deletion mutation was prevalent and may represent an established molecular signature of a circulating HAdV-B7 sublineage. How this mutation affects viral pathogenicity remains unclear and needs further investigation. Our findings underscore the critical importance of implementing timely molecular surveillance for guiding outbreak control and informing prevention strategies against respiratory pathogens.

During the early 2025/26 influenza season, influenza A(H3N2) subclade K rapidly predominated in Beijing, China. Using a test-negative design, we estimated influenza vaccine effectiveness (VE) among influenza-like illness outpatients tested between September and December 2025. Among 9,579 participants, sequencing of 316 randomly selected A(H3N2)-positive samples showed 84.8% were subclade K, and antigenic analysis of 65 viruses indicated antigenic divergence. Despite this, adjusted VE against laboratory-confirmed influenza was 41.3% (95% CI: 29.2 to 51.3), indicating moderate protection during this subclade K-dominated season.

We estimated early influenza vaccine effectiveness (VE) for the 2024/25 season in outpatients, in Beijing using a test-negative design. A(H1N1)pdm09 dominated (99.3%), all sequenced strains (n = 38) clustered in clade 6B.1A.5a.2a, and 37 of 38 antigenically similar to the vaccine strain. VE against any influenza virus infection was 48.5% (95% CI: 34.8–59.5) and 48.7% (95% CI: 35.1–59.7) against A(H1N1)pdm09. Vaccination in the current or previous season against any influenza showed a VE of 52.5% to 54.9%, compared to no vaccination in both seasons.

This study aimed to estimate the end-of-season influenza vaccine effectiveness (VE) for the 2024/25 season in Beijing, China. Methods: We used a test-negative design (TND) to assess influenza VE among outpatients with influenza-like illness (ILI) enrolled through the influenza virological surveillance in sentinel hospitals in Beijing from week 44, 2024 to week 14, 2025. Cases were ILI patients who tested positive for influenza; controls were those who tested negative. Results: Among 18,405 ILI patients tested, 3690 (20.0%) were positive for influenza, with A(H1N1)pdm09 as the predominant strain (98.9%). The overall influenza vaccination coverage was 12.4%. Adjusted VE was 48.3% (95%CI: 40.4%–55.3%) against any influenza and 48.2% (95%CI: 40.3%–55.1%) against A(H1N1)pdm09, with the highest VE observed in adults aged 18–59 years (79.0%). The adjusted VE was similar for those vaccinated in 2023/24 only (53.1%) or both 2023/24 and 2024/25 seasons (50.8%), but lower for those vaccinated only in the 2024/25 season (48.5%). The adjusted VE was higher during the epidemic period (52.5%) than in the pre-epidemic (48.1%) and post-epidemic (35.3%) periods. Conclusions: Our findings indicate moderate VE against laboratory-confirmed influenza, especially A(H1N1)pdm09, during the end of the 2024/25 season in Beijing, China. Influenza vaccination provided protective effects across different epidemic periods. These timely estimates support ongoing public health communication and immunization strategies.

What is already known about this topic?:The H3N8 avian influenza virus (AIV) demonstrates considerable capacity for interspecies transmission and has been documented in multiple mammalian hosts, including equine and canine species. During 2022-2023, three laboratory-confirmed human infections with H3N8 were reported in China, heightening public health concerns about the zoonotic spillover potential of H3 subtype AIVs. What is added by this report?:This study reports the isolation of a genetically reassorted, low-pathogenicity H3N8 avian influenza virus (AIV) from an islet in Niukouyu Wetland Park, Beijing Municipality - the first detection of this viral strain in a wild environment within the city. Throat swabs collected from park staff tested negative for influenza viruses. Phylogenetic analysis demonstrated that the viral hemagglutinin gene originated from the Eurasian lineage, while the neuraminidase gene was derived from the North American lineage. Although no direct evidence of human infection has been documented, multiple mutations identified in the virus's internal genes are associated with enhanced replication capacity, increased virulence, and improved adaptation to mammalian hosts. These molecular features indicate a potential risk for cross-species transmission to humans. What are the implications for public health practice?:Given the potential threat that H3N8 AIVs pose to mammalian species, including humans, this study emphasizes the critical need to strengthen influenza surveillance networks and broaden monitoring efforts specifically targeting H3 subtype AIVs.

Background: During the 2022–2023 influenza season, the influenza activities in most regions of China were postponed, including Beijing. The unusually delayed influenza epidemic posed a challenge to the effectiveness of the influenza vaccine. Methods: Using the test-negative design, we evaluated influenza vaccine effectiveness (VE) during the 2022–2023 influenza season against influenza A-associated outpatient and emergency-department-attended influenza-like illness (ILI) in Beijing, China, from 9 January to 30 April 2023. Results: The analysis included 8301 medically attended ILI patients, of which 1342 (46.2%) had influenza A(H1N1)pdm09, 1554 (53.4%) had influenza A(H3N2), and 11 (0.4%) had co-infection of the two viruses. VE against influenza A-associated ILI patients was 23.2% (95% CI: −6.5% to 44.6%) overall, and 23.1%, 9.9%, and 33.8% among children aged 6 months to 17 years, adults aged 18–59 years, and adults aged ≥60 years, respectively. VE against influenza A(H1N1)pdm09 and against influenza A(H3N2) were 36.2% (95% CI: −1.9% to 60.1%) and 9.5% (95% CI: −34.1% to 39.0%), respectively. VE of the group with vaccination intervals of 14–90 days (70.1%, 95% CI: −145.4 to 96.4) was higher than that of the groups with a vaccination interval of 90–149 days (18.7%, 95% CI: −42.4% to 53.6%) and ≥150 days (21.2%, 95% CI: −18.8% to 47.7%). Conclusions: A moderate VE against influenza A(H1N1)pdm09 and a low VE against influenza A(H3N2) were observed in Beijing during the 2022–2023 influenza season, a season characterized with a delayed and high-intensity influenza epidemic. VE appears to be better within three months after vaccination. Our findings indicate a potential need for the optimization of vaccination policies and underscore the importance of continuous monitoring of influenza to enhance vaccines and optimizing vaccination timing.

雷东多病毒是新发现的一种环状单链DNA病毒，分为Vientovirus和Brisavirus两个种，在人呼吸道中普遍存在，可引起呼吸道系统疾病。本文对雷东多病毒的基因组特征、致病性、流行病学、实验室检测等方面进行综述。

Objective:To characterize the epidemic of influenza in Beijing from 2022 to 2023 and the variation of gene and antigenicity of hemagglutinin (HA) of influenza A H3N2 virus, so as to provide scientific basis for influenza prevention and control in Beijing.Methods:Statistical analysis was carried out on the result of influenza pathogenic monitoring in Beijing from week 14, 2022 to week 20, 2023, and 79 strains of influenza A H3N2 virus were selected at different time and population sources, and their genetic variation and evolution characteristics were analyzed through HA gene amplification sequencing and antigenicity analysis.Results:From week 14, 2022 to week 20, 2023, 24 244 throat swabs of influenza like cases were collected in Beijing, and 4 987 influenza virus nucleic acid positive cases were detected, including 2 749 influenza A H3N2 positive cases, with a detection rate of 11.34%. Among the 79 strains, 50 strains (63.29%) showed low response, 94.44% of the strains from August to November 2022 had low response, and 54.10% of the strains from February to March 2023 had low response, with a statistically significant difference ( χ2=8.079, P=0.004). Compared with the vaccine strain A/Darwin/9/2021, the HA gene sequence of 79 strains of influenza A H3N2 showed nucleotide similarity of 97.47% to 98.47% and amino acid similarity of 97.05% to 98.17%. Genetic evolution analysis showed that the 18 strains isolated from August to November 2022 were all distributed in the 3C.2a1b.2a.1a.1 branch, while the 61 strains isolated from February to March 2023 all belonged to the 3C.2a1b.2a.3a.1 branch. Compared with the vaccine strain, there were multiple site mutations distributed at multiple antigenic determinants and receptor binding sites in A, B, C, D, and E. All strains had potential glycosylation sites of 8NST, 22NGT, 38NAT, 45NSS, 63NCT, 126NWT, 133NGT, 246NST, 285NGS, 483NET, while one strain missed 165NVT glycosylation sites; 55 strains between February and March 2023 missed 122NES glycosylation sites. Conclusions:The HA gene locus of influenza A H3N2 virus detected in Beijing from week 14, 2022 to week 20, 2023 showed multiple mutations, continuous monitoring of this subtype variation is crucial.

Objective:To understand the epidemic situation of Redondoviridae in Beijing and analyze its epidemiologic characteristics.Methods:Pharyngeal swab samples of healthy people and patients with acute respiratory infection in Beijing, including influenza like cases and severe acute respiratory infection (SARI) cases in hospitals were collected. Real time PCR was used to detect the nucleic acid of Redondoviridae. The positive samples were amplified and sequenced to analyze their species. The age and sex distribution of patients and species distribution of Redondoviridae were obtained through statistical analysis. Multiplex PCR was used to detect other common respiratory pathogens in the positive samples of Redondoviridae in influenza like cases and SARI cases, and the pathogenicity of Redondoviridae was analyzed.Results:The positive rates of Redondoviridae in healthy people and acute respiratory infection cases were 20.48% (189/923) and 11.23% (43/390), respectively, with a statistically significant difference ( P<0.05). The positive rate of male was higher than that of female in the healthy population, and the positive rate of the elderly group was higher than that of the adult group and the underage group, with a statistically significant difference ( P<0.05). The positive rate of male patients with acute respiratory tract infection was higher than that of female patients, but there was no significant difference. The proportion of Vientovirus in the positive samples of Redondoviridae was higher than that of Brisavirus, and the difference was statistically significant ( P<0.05). Among the throat swabs of respiratory tract infection cases, 43 were positive for Redondoviridae, of whom 24 were not detected for other pathogens. Conclusions:Redondoviridae widely exists in healthy people of all age groups in Beijing, and is also found in acute respiratory infection cases. The positive rate of Redondoviridae is different in different ages and genders. Both Vientovirus and Brisavirus were detected, and the proportion of Vientovirus was significantly higher than Brisavirus.

Objective:To investigate the phylogenetic and antigenic characteristics of hemagglutinin (HA) gene of influenza B/Victoria lineage (BV) viruses in Beijing during the 2021-2022 influenza surveillance season, and to analyze whether the circulating BV viruses match the vaccine strain.Methods:Pharyngeal swab specimens from influenza like-illness (ILI) cases in the 2021-2022 influenza surveillance season were collected from surveillance network labs in Beijing and cultured in MDCK cells and chicken embryo to isolate BV viruses. Nucleic acids of the viruses were extracted, and the HA gene was amplified and sequenced. The nucleotide and amino acid sequence identity of the HA gene was analyzed using MEGA5.0 software. A phylogenetic tree of HA gene was constructed using the maximum likelihood method. The N-glycosylation sites in HA were predicted online. Three-dimensional structure of HA was constructed using SWISS-MODEL homologous modeling. Hemagglutination inhibition (HI) test was performed to analyze the antigenicity of BV viruses.Results:A total of 402 BV viruses were collected and 58 strains with full-length HA gene sequences were chosen for further analysis. Compared with the HA gene of this year′s vaccine strain (B/Washington/02/2019), there were 27 amino acid mutations, 11 of which were located in four different antigenic determinants. The phylogenetic analysis revealed that three subgroups of 1A.3, 1A.3a1, and 1A.3a2 co-circulated in Beijing with 54 strains (54/58, 93.10%) clustered to the Clade 1A.3a2, two strains (2/58, 3.45%) clustered to the Clade 1A.3a1, and two strains (2/58, 3.45%) in the same subgroup (Clade 1A.3) as the vaccine component BV strain in 2021-2022. Compared with the vaccine strain (B/Washington/02/2019), two BV strains had an additional N-glycosylation site at residue 197, while the other 56 strains showed no change in N-glycosylation sites. Antigenic analysis showed that 35 BV strains (35/58, 60.34%) were antigenically similar to the vaccine strain and 23 strains (23/58, 39.66%) were low-response strains.Conclusions:Three subgroups of BV viruses co-circulated in Beijing during the 2021-2022 influenza surveillance season. The predominant subgroup was Clade 1A.3a2 (93.10%), showing a certain genetic distance with the vaccine strain (B/Washington/02/2019). Nearly 40% (39.66%) of the viruses were low-response strains. This study indicated that continuous monitoring of the variations of influenza epidemic strains and timely providing laboratory basis for screening vaccine component strains were the basic technical guarantee for coping with influenza pandemic.

Objective To identify the pathogen and track the genetic source of a cluster of cases with fever in a kindergarten in Fengtai district during the normalization of COVID-19 prevention and control in Beijing.Methods A descriptive analysis method was used to investigate this cluster of cases with fever in April 2021.Pharyngeal swabs were collected and viral nucleic acid was extracted, real-time PCR was performed to identify SARS-CoV-2 and other common respiratory virus. G gene of human metapneumovirus（hMPV） was amplified by RT-PCR and was then sequenced. BioEdit was used for G gene sequence analysis and the Neighbor-Joining model in MEGA 5. 0 software was used to construct the phylogenic tree of G gene. Results A total of 16 cases were reported in one class with the incidence of 53. 3%（16/30） during 8 days of a cluster outbreak. All pharyngeal swabs collected from 12 cases were tested SARS-CoV-2 negative, six were found to be hMPV positive by multiplex-PCR, and one was positive for both human adenovirus and hMPV. Full-length sequences of G genes were obtained from 2 strains of hMPV. Sequence analysis showed that both strains were hMPV B2 and the nucleic acid homology of G gene was 96. 73%-98. 01% with strains from Japan（LC337940, LC337935, LC1922349） in 2016 and over 98. 40%with strains from Shandong（OL625642, OL625644） in 2019, Henan MN944096 in 2019.Compared with the amino acid sequence of hMPV-B2 reference strain（AY297748）, six amino acid insertions containing EKEKEK were identified between 161-166 amino acid location and N-glycosylation of G protein analysis showed that the two strains had four N-glycosylation sites. Conclusions The leading pathogen for this cluster outbreak is found to be hMPV-B2, which are highly homologous with strains from Japan, Shandong and Henan. Therefore, a non-stop surveillance of hMPV is necessary during the normalization control and prevention period for COVID-19.

Viral isolation in cell cultures has been regarded for decades as the “gold standard” for the laboratory diagnosis of influenza viral infections. Not all viral strains could be isolated from clinical samples. This study aimed to quantify the viral load in the samples before isolation to save working time and improve working efficiency. Four hundred samples from patients with influenza-like cases were confirmed pdmH1N1 positive (200 cases) and B Victoria (BV) positive (200 cases) by whole-genome sequencing and analyzed by ddPCR for viral load in samples before isolation, and isolation results were verified by hemagglutination (HA) assay and hemagglutination-inhibition (HI) tests. Probit regression analysis was used to calculate the isolation viral load limit with a 95% probability level by SPSS 19.0 software. The results showed that the isolation limit of viral load was 4.9 × 104 (95% CI: 2.5 × 104–9.0 × 104) copies/mL for pdmH1N1 and 1.9 × 104 (95% CI: 7.8 × 103–3.6 × 104) copies/mL for BV. The isolation rate of clinical samples is positively correlated with the viral load in clinical samples, which can be used for viral culture, providing important guidance for daily work.

Objective:To analyze genetic and phylogenic characteristics of hemagglutinin (HA) gene of the first outbreak of influenza B Victoria virus (BV) in Beijing during 2021-2022 influenza epidemic.Methods:Pharyngeal swabs from influenza-like illnesses of the first influenza B outbreak in Beijing were collected. After extracting nucleic acid, the next generation sequencing (NGS) technology was used for sequencing and analysis. The Neighbor-Joining model of Mega 6.0 software was used to construct the phylogenic tree of HA gene, and the nucleotide and amino acid sequence identity were conducted.Results:Full-length sequence of two HA gene of BV strains were obtained by NGS. Compared with the HA gene of this influenza season vaccine strain (B/Washington/02/2019), there were nine amino acid mutations, six of which were located in three different antigenic determinants. Furthermore, the phylogenetic tree analysis showed that the HA gene of this outbreak were all located in Clade 1A.3a2 branch, not in the same strain of influenza vaccine recommended in the northern hemisphere 2021-2022 (B/Washington/02/2019).Conclusions:HA gene of this BV epidemic strain has many variations in antigenic epitopes. It is necessary to strengthen the surveillance and analysis of variation regularity of BV influenza, to provide a strong data support for BV prevention and control strategy for BV prevention and control strategy .

Objective:To understand the characteristics of hemagglutinin (HA) gene and antigenicity variation of influenza B Victoria lineage virus (BV) in Beijing in 2021, so as to provide scientific basis for influenza prevention and control in Beijing.Methods:The etiological surveillance results of influenza in Beijing from January to November 2021 were analyzed with statistical methods. Fifteen BV strains were randomly selected. The HA genes were amplified and sequenced. To analyze the gene mutation and evolutionary characters by hemagglutinin gene amplification and sequencing.Results:From January to November 2021, 16 097 samples of influenza like cases were collected. Two hundred and eight samples were positive by nucleic acid test. Two hundred and five samples were identified as BV (98.56%), and 3 samples were influenza B Yamagata lineage virus (1.44%). The similarity of nucleic acid and amino acid sequences between the 15 BV stains and the vaccine strain B/Washington/02/2019 were 98.39% - 98.75% and 97.86% - 98.40%, respectively. Phylogenetic analysis indicated that the 15 BV stains belonged to Clade1A.3a2. Multiple point mutations existed compared to the vaccine stain. There were 11 glycosylation sites in the 15 BV strains. One strains (6.6%) was a low response strain of the vaccine strain.Conclusions:BV strains were predominant among the influenza viruses observed in Beijing in 2021. HA gene loci showed multiple mutations, suggesting the importance of continuous surveillance of the mutations in this subtype for providing a scientific basis for influenza prevention and control strategies formulation.

Coronavirus disease 2019 (COVID-19) has spread widely around the world, and in-depth research on COVID-19 is necessary for biomarkers and target drug discovery. This analysis collected serum from six COVID-19-infected patients and six healthy people. The protein changes in the infected and healthy control serum samples were evaluated by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and high-performance liquid chromatography (HPLC). The differential protein signature in both groups was retrieved and analyzed by the Kyoto Encyclopedia of Gene and Genomes (KEGG), Gene ontology, COG/KOG, protein–protein interaction, and protein domain interactions tools. We shortlisted 24 differentially expressed proteins between both groups. Ten genes were significantly up-regulated in the infection group, and fourteen genes were significantly down-regulated. The GO and KEGG pathway enrichment analysis suggested that the chromosomal part and chromosome were the most enriched items. The oxytocin signaling pathway was the most enriched item of KEGG analysis. The netrin module (non-TIMP type) was the most enriched protein domain in this study. Functional analysis of S100A9, PIGR, C4B, IL-6R, IGLV3-19, IGLV3-1, and IGLV5-45 revealed that SARS-CoV-2 was closely related to immune response.

Since 2010 the year when it was first reported in domestic ducks in China, highly pathogenic avian influenza (HPAI) H5N8 has caused several outbreaks in different countries. The first outbreak wave was documented in South Korea and Japan in 2014 and the second wave was reported in Asian and European countries in 2016. More importantly, zoonotic infection was first reported in poultry workers in Russia in 2021. Therefore, active surveillance on H5N8 is highly needed. Surveillance on live birds instead of environmental samples is commonly reported. In the present study, we reported detection and genomic characterization of an environmental H5N8 strain in environmental samples of Tongzhou poultry meat markets in Beijing on a monthly basis from March 2021 to February 2022. Among 600 samples screened, a total of 27 samples were positive for influenza A virus with 4 typed as H5N8, 10 H7N9, and 13 H9N2. Whole genome sequencing and analysis of one duck neck with a higher virus load showed that A/Environment sample/Beijing/TZ001/20 21 (H5N8) clade 2.3.4.4b had the highest identities (over 99%) in all eight segments with H5N8 isolates from wild birds swan and tern in Hubei and had polybasic cleavage site PLREKRRKR/G, characteristic of a HPAI virus. Overall, our data indicate that HPAI H5N8 virus is still circulating in domestic ducks in China in the study period and continued surveillance in domestic and wild birds is needed to control H5N8.

Objective:To establish a nested PCR method to detect the 2019 novel coronavirus (2019-nCoV), as a supplement to the real-time fluorescent PCR method, and discuss the preliminary application value of this method in clinical diagnosis.Methods:According to the conservative sequences of the 2019-nCoV gene, the nested PCR primers including N gene and S gene, were designed on line. By optimizing the nested PCR reaction systems, the qualitative detection was established by testing N gene and sequencing its PCR product while the preliminary type identification was established by testing S gene and sequencing its PCR product. The sensitivity was evaluated by the gradient dilution of 2019-nCoV positive samples’ nucleic acid and the specificity was evaluated by detecting the human coronavirus OC43, 229E, HKU1, NL63, influenza virus positive samples. The established method was applied to 15 samples with Ct >33 and 15 samples with Ct <33 screened by real-time fluorescent PCR, and the positive amplification result were sequenced and analyzed to verify the result. Results:The established nested PCR method could amplify specific bands of 355 bp N gene fragment and 449 bp S gene fragment. No amplifications occurred in other human coronaviruses samples including 229E、OC43、HKU1、NL63 or in influenza virus samples including H3N2, H1N1(pdm) and B. The minimum detection limit of the N gene fragment could reach Ct value about 37.21. Among the 30 COVID-19 positive samples, the N gene positive coincidence rate detected by nested PCR was 100% (30/30); the S gene positive coincidence rate reached 60% (18/30). 28 samples’ sequences of N gene fragment were completely consistent with 2019-nCoV by BLAST, and the characteristic result of site mutations of 12 samples’ S gene was obtained. Conclusions:A nested PCR method for the specific detection of 2019-nCoV was established, and some characteristic mutations on S gene could be analyzed by sequencing the PCR amplified products. It could be used as a supplement to the real-time fluorescent PCR method.

Objective:To clarify the M protein ( emm gene) types and drug susceptibility characteristic variations of Group A Streptococcus (GAS) in children in Beijing. Methods:The GAS strains isolated from throat swab samples of children diagnosed with scarlet fever and pharyngeal infection in scarlet fever etiology surveillance sentinel hospitals in 16 districts of Beijing in 2018, 2019 and 2021 were analyzed retrospectively.PCR amplification and sequencing were used for emm genotyping, and the minimum inhibitory concentrations (MIC) of 10 antibiotics were determined by the broth microdilution method.The data were analyzed using χ2 test and Fisher′ s exact method between groups. Results:A total of 557 GAS strains were collected, and 11 emm genotypes ( emm1, emm3, emm4, emm6, emm11, emm12, emm22, emm75, emm89, emm128, and emm212) were detected.Of 557 strains, 238 trains were of emm1 type (42.73%), 271 strains were of emm12 type (48.65%) and 48 strains were of other emm types (8.62%). The detection rates of emm1, emm12 and other emm type genes in 2018, 2019, and 2021 were [37.50% (105/280 strains), 57.14% (160/280 strains), 5.36% (15/280 strains)], [49.05% (129/263 strains), 39.54% (104/263 strains), 11.41% (30/263 strains)], and [28.57% (4/14 strains), 50.00% (7/14 strains), 21.43% (3/14 strains)], respectively.In children infected with emm12 in 2018 and 2019, there were more children under 6 years old than children over 6 years old (62.50% vs.46.88%, 46.36% vs.30.36%) (χ 2=7.182, 6.973; all P<0.05). Drug susceptibility testing results suggested that 225 randomly selected GAS strains were all 100.00% sensitive to 7 antibiotics including Penicillin, Levofloxacin, Meropenem, Linezolid, Cefotaxime, Cefepime and Vancomycin.The rates of resistance to Erythromycin, Tetracycline and Clindamycin were [88.57% (93/105 strains), 87.62% (92/105 strains), 86.67% (91/105 strains)], and [94.34% (100/106 strains), 94.34% (100/106 strains), 87.74% (93/106 strains)] in 2018 and 2019, respectively.The test strains were 100.00% (14/14 strains) resistant to the above 3 antibiotics in 2021.MIC 50 and MIC 90 values of Penicillin in 2018, 2019, and 2021 were (0.03 mg/L, 0.03 mg/L), (0.03 mg/L, 0.06 mg/L), and (0.06 mg/L, 0.06 mg/L), respectively.Among 225 GAS strains, 207 strains had drug resistance and were resistant to more than one drug.Specifically, 94.69% (196/207 strains) were resistant to Erythromycin, Tetracycline and Clindamycin.About 4.35% (9/207 strains) were resistant to both Erythromycin and Clindamycin.A total of 0.97% (2/207 strains) were resistant to Erythromycin and Tetracycline. Conclusions:The emm genotypes of GAS in children in Beijing are diverse in 2018, 2019 and 2021.The dominant genotypes are emm12 and emm1, and emm12 is the main epidemiological type.GAS strains maintain highly resistant to Erythromycin, Clindamycin and Tetracycline, and sensitive to Penicillin and other antibiotics.However, MIC 50 and MIC 90 of Penicillin shows an ascending trend.