ABSTRACT Human adenovirus (HAdV)−114 (P7H3F3) is a newly identified recombinant genotype, while recent studies suggest that it likely represents a previously circulating HAdV‐3 strain. A total of 413 strains previously identified as HAdV‐3 from 20 Chinese provinces during 2000–2024, along with 125 GenBank‐derived HAdV‐3 strains (1988–2023), underwent genotype reconfirmation and genetic evolutionary analysis using the penton base, hexon, and fiber genes. Furthermore, 97 representative strains were selected for whole‐genome analysis to determine their genetic recombination patterns. Of the 538 analyzed strains, 537 were identified as HAdV‐114 using three genes, with only one 2010 U.S strain classified as HAdV‐3. Phylogenetic analysis revealed three HAdV‐114 clades (Clades 1–3): Clade 1 (1988–2019) displayed sporadic circulation, Clade 2 (2023–2024) appeared geographically restricted to three Chinese provinces, and Clade 3 (2000–2024) likely represented the globally dominant strain. Genetic evolutionary analyses indicated that, although HAdV‐114 originated from recombination between HAdV‐7 and HAdV‐3, its three Clades diverged from the parental trajectories, with Clades 1 and 3 sharing a common evolutionary origin and Clade 2 forming an independent terminal cluster. Genomic recombination analysis identified two recombination patterns across the three clades. All clades shared a 3′ terminal genomic region (nt 18,794–end) derived from HAdV‐3, whereas their 5′ ends comprised gene fragments recombined from unknown sources and epidemic HAdV‐7 strains, with varied breakpoints. HAdV‐114, historically identified as HAdV‐3, is the predominant genotype associated with HAdV‐related respiratory infections. Therefore, enhanced surveillance of recombinant HAdV strains is imperative for effective disease control.
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