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目的:了解2020—2022年青岛市手足口病病原学特征和柯萨奇病毒A6型(coxsackievirus A6，CVA6)的VP1基因进化特征，为手足口病的防控工作提供依据。方法:利用荧光RT-PCR对青岛地区2020—2022年的手足口病患者咽拭子样本进行总肠道病毒(EV)、肠道病毒71型(EV71)、CVA16、CVA6和CVA10检测，并对CVA6阳性分离株进行VP1基因全长序列扩增及基因序列测定，以MEGA-X软件进行系统发育及分子特征分析。结果:1 645份手足口病患者样本中肠道病毒总阳性率为80.73%(1 328/1 645)。EV71、CVA16、CVA6、CVA10及其他肠道病毒构成比分别为0.08%(1/1 328)、19.65%(261/1 328)、68.15%(905/1 328)、0.98%(13/1 328)、11.14%(148/1 328)，5岁以下儿童占全部病例的75.68%(1 245/1 645)，占重症病例的100%(2/2)。获得的45株CVA6 VP1区基因测序绘制了基因树，属于D3a亚型，基因进化分析其VP1区蛋白质发生了10处氨基酸的非同义突变。结论:2020—2022年青岛地区手足口病以CVA6 D3a亚型为主要流行株，且在5岁以下高发。

Objective:To investigate the etiological characteristics of hand, foot and mouth disease (HFMD) and the VP1 gene evolution of coxsackievirus A6 (CVA6) in Qingdao from 2020 to 2022.Methods:Total enterovirus (EV), enterovirus 71 (EV71), coxsackievirus A16 (CVA16), CVA6 and coxsackievirus A10 (CVA10) were detected by fluorescent RT-PCR in throat swab samples from HFMD patients in Qingdao from 2020 to 2022. The full-length sequence amplification of VP1 gene and gene sequence determination of CVA6 isolates were performed, and the phylogenetic and molecular characteristics were analyzed by MEGA-X software.Results:The overall positive rate of enteroviruses in 1 645 samples collected during 2020-2021 was 80.73%(1 328/1 645). The proportions of EV71, CVA16, CVA6, CVA10 and other enteroviruses were 0.08%(1/1 328), 19.65%(261/1 328), 68.15%(905/1 328), 0.98%(13/1 328) and 11.14%(148/1 328), respectively. Children under 5 years old accounted for 75.68% (1 245/1 645) of all cases and 100% (2/2) of severe cases. A phylogenetic tree was constructed based on the VP1 region sequences of 45 CVA6 strains showing all strains belonging to the D3a subtype. There were 10 non-synonymous mutations of amino acids in the VP1 protein by gene evolution analysis.Conclusions:CVA6 D3a subtype is the main epidemic strain causing HFMD in Qingdao from 2020 to 2022, especially in children under 5 years old.