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分析2023年广东省急性出血性结膜炎（Acute hemorrhagic conjunctivitis,AHC）疫情流行末期病原体组成及其分子特征情况，为AHC的防治提供科学依据。本研究采集AHC流行末期81份AHC患者眼结膜拭子，采用实时荧光定量PCR(Real-time quantitative PCR,qPCR)方法，对其进行人肠道病毒70型（Human enterovirus 70,HEV70）、柯萨奇病毒A组24型变异株（Coxsackievirus A24 variant,CVA24v）和腺病毒（Adenovirus,AdV）核酸检测。通过HEp-2细胞进行病毒分离。对分离株进行PCR扩增和基因测序。用Mega11和BioEdit软件进行基因进化分析，用SWISS-MODEL进行VP1蛋白三维结构分析，通过IEDB进行VP1蛋白的B细胞抗原表位预测。结果显示81份眼拭子样本荧光定量PCR检测，EV70全部阴性，20份样本CVA24v阳性，16份样本AdV阳性。使用HEp-2细胞成功分离到8株CVA24v毒株。基于分子测序分析发现，2023年AHC流行末期的CVA24v病毒株与5月份疫情初期病毒株相比，8株CVA24v毒株在3C区核苷酸共有7处突变，其中在第309位点共享同义突变C→T。在VP1区共有19处突变，除AHC705/GD/CHN/2023株外，其余7株在第30位点共享同义突变T→C。此外还获得一株CVA24v的全基因组序列，和流行初期病毒株全长序列相比有24个核苷酸突变，3个氨基酸突变。本研究通过分析VP1蛋白的三维结构预测，识别出了本次疫情流行末期相比早期出现三个变异位点，其中T98A和A146T两个位点位于蛋白表面，参与组成抗原决定簇。研究结果显示，与疫情初期由CVA24v独立引起的流行相比，流行末期出现了CVA24v与AdV共同流行的现象。这一发现对理解病毒变异与流行动态具有重要意义。
Abstract：In this paper, the pathogen composition and molecular characteristics at the end of the acute hemorrhagic conjunctivitis(AHC) epidemic in Guangdong Province,China in 2023 were analyzed, in attempts to provide scientific basis for the prevention and treatment of AHC. In this study, a total of 81 conjunctival swab samples were collected from AHC patients, and nucleic acid detection for human enterovirus 70(HEV70), coxsackievirus A24 variant(CVA24v) and adenovirus(AdV) was performed with real-time quantitative PCR(qPCR). The viruses were isolated using HEp-2 cells, followed by PCR amplification and gene sequencing.Genetic evolutionary analysis was conducted using Mega11 and BioEdit software, the three-dimensional structure of VP1 protein was analyzed through SWISS-MODEL, and the B cell epitopes of VP1 protein were examined using IEDB. The qPCR results show that in the 81 eye swab samples, there are 81 negative EV70samples, 20 positive CVA24v samples and 16 positive AdV samples. were used to 8 CVA24v strains were successfully isolated using HEp-2 cells. Molecular sequencing analysis shows 8 CVA24v strains at the end stage of the 2023 AHC epidemic have 7 nucleotide mutations in the 3C zone, including a shared synonymous mutation C→T at site 309. There are 19 mutations in the VP1 zone, where 7 strains share a synonymous mutation T→C at site 30. Furthermore, a full-genome sequence of a CVA24v strain was obtained and found to have 24nucleotide and 3 amino acid mutations, which were different from the virus strains at the early stage of the epidemic. In this study, three new mutation sites were identified at the end of the current epidemic by analyzing the 3D structure of the VP1 protein, of which T98A and A146T sites are located on the protein surface and are involved in forming the antigenic determinant cluster. The results imply a co-epidemic of CVA24v and AdV at the end stage of the epidemic, in contrast to the early stage dominated by CVA24v. This discovery is crucial for understanding the viral variation and epidemic dynamics.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.004583
中图分类号:R777.31
引用信息:
[1]杨银柯,钟干芳,徐俊贤等.2023年广东省急性出血性结膜炎流行末期病原组成及基因进化分析[J].病毒学报,2024,40(05):1010-1021.DOI:10.13242/j.cnki.bingduxuebao.004583.
基金信息: