A preliminary study of Jingmen tick virus infection in human and domestic animals in Hubei, 2022
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摘要:
目的 对2022年湖北省人和家养动物血清中荆门蜱病毒（JMTV）的抗体阳性率进行初步研究，评估荆门蜱病毒感染人群和动物的风险。 方法 将JMTV NS3蛋白在大肠埃希菌中表达并纯化，制备ELISA包被抗原，利用人和动物血清分别确定cut-off值；收集湖北多市牛羊养殖场采集的牛、羊血清，武汉市中心医院体检健康人血清，对这些血清样本以及实验室保存的林区居民血清进行JMTV特异性抗体以及核酸检测。 结果 初步建立基于NS3蛋白的JMTV 抗体检测ELISA方法。 确定人血清、牛血清和羊血清的cut-off值分别为0.342、0.098和0.298。 44份牛血清和193份羊血清中JMTV抗体阳性率分别为4.55%和3.63%。 174份体检健康人和13份林区居民血清JMTV抗体结果均为阴性。 所有样本中均未检测到JMTV的核酸片段。 结论 家养动物牛、羊血清中检出JMTV特异性抗体，提示需关注牛、羊作为蜱携带者导致病毒扩散的风险。 Abstract:Objective T o study the positive rate of Jingmen tick virus (JMTV) antibody in serum of domestic animals and assess the risk of JMTV infection in humans and domestic animals in Hubei province, 2022. Methods JMTV NS3 protein as antigen was expressed in Escherichia coli and purified. ELISA envelope antigen was prepared, and cut-off values were determined by human and animal serum. Serum samples from cattle, sheep from farms and serum samples from healthy people from hospitals were collected in Hubei. JMTV-specific antibody and nucleic acid were detected in theses samples, as well as serum from forest residents stored in our laboratory. In addition, JMTV-specific nucleic acid was detected in all samples by RT-PCR method. Results An ELISA assay for JMTV antibody detection based on NS3 protein was established. The cut-off values of human, bovine and sheep serum were determined to be 0.342, 0.098 and 0.298, respectively. The positive rates of JMTV antibody in 44 bovine and 193 sheep serum were 4.55% and 3.63%, respectively. JMTV-specific antibody were negative in 174 healthy subjects and 13 forest residents. No nucleic acid fragments of JMTV were detected in all samples. Conclusion JMTV specific antibody was detected in the serum of domestic cattle and sheep, suggesting that it is necessary to pay attention to the spread risk of the virus caused by cattle and sheep. -
Key words:
- Jingmen tick virus /
- NS3 /
- Enzyme-linked immunosorbent assay /
- Serological screening
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图 1 NS3信号肽分析
注：Sec/SPI. 由 Sec 转座转运，并由信号肽酶 I (Lep) 切割的“标准”分泌信号肽；CS. 用来区分是否为剪切位点，最高峰值为剪切位点后的第一个氨基酸(即成熟蛋白的第一个氨基酸残基)；S-score. 用来区分相应位置是否为信号肽区域
Figure 1. NS3 Signal peptide analysis
图 3 NS3蛋白小量诱导表达及表达形式的SDS-PAGE鉴定
注：a. NS3蛋白小量诱导表达SDS-PAGE鉴定：M. Marker；泳道1. 未诱导；泳道2～4. 1 mmol/LIPTG诱导不同单克隆菌株。b. NS3蛋白表达形式SDS-PAGE鉴定：M. Marker；泳道1. 上清；泳道2. 沉淀
Figure 3. SDS-PAGE identification of small amount induced expression and expression form of NS3 protein
图 4 NS3蛋白表达（包涵体）纯化SDS-PAGE结果
注：M. Marker；泳道1. 过柱后；泳道2～6. 300 mmol/L咪唑洗脱的NS3重组蛋白
Figure 4. SDS-PAGE identification of NS3 protein expression (inclusion bodies) purification
图 5 NS3蛋白his单抗Western Blot分析
注：M. Marker；泳道1. NS3蛋白
Figure 5. Western blot analysis of NS3 protein his monoclonal antibod
图 6 人群血清样本A450值分布
注：A450. 吸光值
Figure 6. Distribution of A450 value in serum samples of people
图 7 动物血清样本A450值分布
注：A450 吸光值
Figure 7. Distribution of A450 value in serum samples of beast
表 1 动物采样信息表
Table 1. Information about animal samples
采样时间 地点 养殖场名称 种属 样本类型 样本数量 2022年5月 湖北宜昌市 CY 羊 血清 50份 XT 羊 血清 8份 YD 羊 血清 10份 DJ 羊 血清 9份 GX 羊 血清 10份 2022年6月 湖北宜昌市 CY 羊 血清（血块） 31份（31份） 湖北黄冈市 MC 羊 血清（血块） 34份（34份） LT 羊 血清（血块） 41份（41份） 湖北十堰市 ZS 牛 血清（血块） 5份（5份） 湖北宜昌市 ZJ 牛 血清（血块） 14份（14份） 湖北襄阳市 ZY 牛 血清（血块） 25份（25份） 
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表 3 不同养殖场血清样本JMTV阳性率（%）
Table 3. Positive rate of JMTV in serum samples from different farms
样本类型 养殖场名称 样本数量 阳性例数 阳性率 牛血清 ZS 5 0 0.00 ZJ 14 1 7.14 ZY 25 1 4.00 羊血清 CY 81 6 7.41 XT 8 0 0.00 YD 10 0 0.00 DJ 9 0 0.00 GX 10 0 0.00 LT 41 1 2.44 MC 34 0 0.00
下载: 导出CSV
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