Abstract：Objective To compare the detection rate of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the temporal changes in the concentrations of different target genes and the composition of variants in sewage in Zhengzhou City and Luoyang City of Henan Province in 2024, so as to provide data references for regional sewage surveillance. Methods From January to December 2024, the main inlet channels of 5 and 4 domestic wastewater treatment plants were selected in the main urban areas of Zhengzhou City and Luoyang City, respectively, for continuous sewage sample collection for 53 weeks. An automatic sewage sampler was used to collect flow-proportional mixed samples within 24 h, and the virus was enriched and concentrated by the polyethylene glycol precipitation method. The N gene and ORF1ab gene were detected by reverse transcription real-time fluorescence quantitative PCR, and the detection rate of SARS-CoV-2 was calculated. A standard curve was prepared with standard substances for quantitative analysis. The Illumina sequencer was used for whole-genome sequencing of positive samples with Ct values ≤34, and the composition of variants was analyzed. Results A total of 690 sewage samples were collected and tested in Zhengzhou City and Luoyang City in 2024, and 477 SARS-CoV-2-positive samples were detected, with a detection rate of 69.13%. The median concentrations of N gene and ORF1ab gene were 3.43 (interquartile range, 7.31) copies/mL and 1.24 (interquartile range, 5.93) copies/mL, respectively. The SARS-CoV-2 detection rate in Zhengzhou City was lower than in Luoyang City (62.82% vs. 75.82%, P<0.05). The median concentrations of N gene and ORF1ab gene in Zhengzhou City were 5.98 (interquartile range, 10.32) copies/mL and 5.16 (interquartile range, 17.33) copies/mL, respectively, which were higher than 2.01 (interquartile range, 3.81) copies/mL and 0.73 (interquartile range, 1.03) copies/mL in Luoyang City (both P<0.05). The SARS-CoV-2 detection rate in both cities showed a temporal trend of a high level in the early stage, fluctuation in the middle stage, and decline in the late stage. The SARS-CoV-2 concentration in Zhengzhou City peaked at weeks 21, 28 and 32, with the maximum ORF1ab gene concentration reaching 242.05 copies/mL. The overall SARS-CoV-2 concentration in Luoyang City was relatively low, with three peaks appearing at weeks 10, 18 and 33. Whole-genome sequencing results showed that Omicron variants and their sublineages were dominant in SARS-CoV-2 lineages from sewage in both cities. Nine sublineages were detected in Zhengzhou City, including JN.1, BA.2, LB.1, LA.1, LA.2, XDV, KP.1, KP.4 and XDP; six sublineages were detected in Luoyang City, including JN.1, EG.5, XBB, XBN, HK.3 and BA.2. The dominant lineage in Zhengzhou City shifted from JN.1 to XDV, KP.1 and other sublineages in the late surveillance period, while JN.1 remained the dominant lineage in Luoyang City. Conclusions There were regional differences in SARS-CoV-2 detection rate, viral concentration and variant composition in sewage between Zhengzhou City and Luoyang City in 2024. Luoyang City had a higher viral detection rate, while Zhengzhou City had higher viral concentration and more active variant turnover.
[1] Zhang Q F,Gao C Z,Ge X Y,et al.Tracking viral variants by wastewater surveillance,from laboratory diagnosis towards on-site monitoring:lessons learned from the SARS-CoV-2 pandemic[J/OL].J Environ Manag,2026,404[2026-09-19].https://doi.org/10.1016/j.jenvman.2026.129353. [2] Munteanu V,Saldana M A,Dreifuss D,et al.SARS-CoV-2 wastewater genomic surveillance:approaches,challenges,and opportunities[J/OL].Genome Biol,2026,27(1)[2026-09-19].https://doi.org/10.1186/s13059-025-03927-6. [3] Alhama J,Maestre J P,Martín MÁ,et al.Monitoring COVID-19 through SARS-CoV-2 quantification in wastewater:progress,challenges and prospects[J].Microb Biotechnol,2022,15(6):1719-1728. [4] Hu B,Guo H,Zhou P,et al.Characteristics of SARS-CoV-2 and COVID-19[J].Nat Rev Microbiol,2021,19(3):141-154. [5] Holmes E C.The emergence and evolution of SARS-CoV-2[J].Annu Rev Virol,2024,11(1):21-42. [6] Ge J B.The COVID-19 pandemic in China:from dynamic zero-COVID to current policy[J].Herz,2023,48(3):226-228. [7] 毋碧聪,宋云,卢世栋,等.河南省2021年境外输入SARS-CoV-2全基因组测序分析[J].郑州大学学报(医学版),2023,58(5):623-627. [8] WS/T 10042—2025 污水中新型冠状病毒监测技术规范[S]. [9] Mannarà G,Martinelli M,Giubbi C,et al.Wastewater surveillance for SARS-CoV-2 in northern Italy:an evaluation of three different gene targets[J/OL].Microorganisms,2025,13(2)[2026-09-19].https://doi.org/10.3390/microorganisms13020236. [10] 董柯,邵束钰,许太彬,等.郑州市城市污水中新型冠状病毒监测结果[J].现代疾病预防控制,2024,35(2):86-90. [11] Zheng X W,Leung K,Xu X Q,et al.Wastewater surveillance provides spatiotemporal SARS-CoV-2 infection dynamics[J].Engineering,2024,40:70-77. [12] Lyu L S,Liu X Y,Sun Q L,et al.Association between SARS-CoV-2 levels in urban wastewater and reported COVID-19 cases in Changsha,Central China[J/OL].BMC Infect Dis,2025,25(1)[2026-09-19].https://doi.org/10.1186/s12879-025-11633-8. [13] Cheng K Y,Lyu Y,Li C K,et al.Meta-analysis of the SARS-CoV-2 positivity rate in municipal wastewater[J/OL].Environ Geochem Health,2024,46(4)[2026-09-19].https://doi.org/10.1007/s10653-024-01895-7. [14] 黄金金,倪德航,徐素芹,等.贵阳市2023年1 034份城市污水新型冠状病毒监测结果分析[J].现代预防医学,2024,51(23):4392-4397. [15] Nappo A,Petricciuolo M,Berno G,et al.One-year monitoring of the evolution of SARS-CoV-2 omicron subvariants through wastewater analysis(central Italy,August2023-July 2024)[J/OL].Life(Basel),2025,15(6)[2026-09-19].https://doi.org/10.3390/life15060850. [16] Veneri C,Brandtner D,Mancini P,et al.Tracking the Spread of the BA.2.86 lineage in Italy through wastewater analysis[J].Food Environ Virol,2024,16(4):449-457. [17] Petala M,Kostoglou M,Karapantsios T,et al.Relating SARS-CoV-2 shedding rate in wastewater to daily positive tests data:a consistent model based approach[J/OL].Sci Total Environ,2022,807(Pt2)[2026-09-19].https://doi.org/10.1016/j.scitotenv.2021.150838. [18] Li X,Zhang S,Shi J,et al.Uncertainties in estimating SARS-CoV-2 prevalence by wastewater-based epidemiology[J/OL].Chem Eng J,2021,415[2026-09-19].https://doi.org/10.1016/j.cej.2021.129039.
