Investigation of an infection case of Orientia tsutsugamushi Boryong sub-genotype in Ningbo, Zhejiang, 2022
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摘要:
目的 对2022年1例恙虫病疑似病例进行实验室检测和分析,了解浙江省宁波市恙虫病东方体基因型流行情况。 方法 对疑似病例进行流行病学调查,询问其病史;采集病例发热期血液,提取总DNA采用实时荧光定量PCR方法扩增目的基因tsa56,根据测得的序列构建系统进化树,确定该恙虫病东方体基因型。 结果 实时荧光定量PCR方法检出病例血液中存在恙虫病东方体;扩增序列经比对后与已知恙虫病东方体Boryong具有最近亲缘关系。 结论 宁波市实验室确诊1例恙虫病东方体感染病例,为Boryong基因亚型。 -
关键词:
- 恙虫病 /
- 恙虫病东方体 /
- Boryong基因亚型 /
- 进化分析
Abstract:Objective To understand the genotype of Orientia tsutsugamushi by analyzing a suspected tsutsugamushi disease case detected in Ningbo, Zhejiang province, 2022. Methods Epidemiological investigation was conducted. Blood samples of the case in fever phase were collected for the extraction of pathogen DNA. Quantitative real-time fluorescent PCR was performed for the amplification of targeted gene tsa56, and a phylogenetic tree was constructed based on gene sequencing result to determine the genotype of O. tsutsugamushi. Results O. tsutsugamushi was detected in the case's blood samples by quantitative real-time fluorescent PCR. Phylogenetic tree analysis demonstrated it had most closely relationship with Boryong strain. Conclusion An infection case of O. tsutsugamushi was laboratory confirmed in Ningbo, the sub-genotype was Boryong. -
表 1 用于扩增恙虫病东方体标志基因tsa56的引物
Table 1. Primers used for amplification of Orientia tsutsugamushi targeted gene tsa56
用途 引物名称 序列(5' ~3') 退火温度(℃) 产物长度( bp) 参考文献 普通PCR方法 nml56kd Ot-F CCTAATAGTGCATCTGTCGAA 51 712 [9] nml56kd Ot-R CTAGAAGTTATAGCGTAC 荧光PCR方法(SYBR染料法) fluo56kd Ot-F AAAGACAAAGAGGCAGAGT 55 212 [9] fluo56kd Ot-R CCTTCAGCAGCATTAATTGC 荧光PCR方法(探针法)a taq56kd-Ot-F TCTRCRCCAGTAATYATTCCTCC 60 130 [10] taq56kd-Ot-R TGTTAATTGCTAGTGCAATGTCTGC taq56kd-Ot-probe FAM-AAGGACCACACTCTAAT-MGB 注:a. 引物序列中R=A/G,Y=T/C -
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