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. -
图 1 2022年宁波市1例恙虫病东方体患者肚脐焦痂
Figure 1. Navel eschar from a tsutsugamushi disease patient in Ningbo in 2022
图 2 实时荧光PCR方法扩增病例血液样品中的目的基因tsa56
注：A. SYBR染料法；B. 探针法
Figure 2. Amplification of targeted tsa56 gene in case’s blood samples by real-time fluorescent PCR
图 3 基于恙虫病东方体部分tsa56基因序列构建的系统发生树
注：本研究恙虫病东方体序列为Orientia tsutsugamushi TXR，其余为参考序列
Figure 3. Phylogenetic tree of Orientia tsutsugamushi based on partial tsa56 gene sequence
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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[1] Prakash JAJ. Scrub typhus: risks, diagnostic issues, and management challenges[J]. Res Rep Trop Med, 2017, 8: 73–83. DOI: 10.2147/RRTM.S105602. [2] Xu G, Walker DH, Jupiter D, et al. A review of the global epidemiology of scrub typhus[J]. PLoS Negl Trop Dis, 2017, 11(11): e0006062. DOI: 10.1371/journal.pntd.0006062. [3] Izzard L, Fuller A, Blacksell SD, et al. Isolation of a novel Orientia species (O. chuto sp. nov. ) from a patient infected in Dubai[J]. J Clin Microbiol, 2010, 48(12): 4404–4409. DOI: 10.1128/JCM.01526−10. [4] Weitzel T, Dittrich S, López J, et al. Endemic scrub typhus in South America[J]. N Engl J Med, 2016, 375(10): 954–961. DOI: 10.1056/nejmoa1603657. [5] Thakur SD. Scrub typhus: an underestimated endemic disease of Northwestern Himalayas, India[J]. Curr Sci, 2021, 121(7): 899. DOI: 10.18520/cs/v121/i7/899−905. [6] Zhang WY, Wang LY, Ding F, et al. Scrub typhus in mainland China, 2006–2012: the need for targeted public health interventions[J]. PLoS Negl Trop Dis, 2013, 7(12): e2493. DOI: 10.1371/journal.pntd.0002493. [7] Fan MY, Walker DH, Yu SR, et al. Epidemiology and ecology of Rickettsial diseases in the People's Republic of China[J]. Rev Infect Dis, 1987, 9(4): 823–840. DOI: 10.1093/clinids/9.4.823. [8] 张萌, 王显军, 赵仲堂. 中国恙虫病流行态势及预防控制[J]. 中华流行病学杂志,2011,32(4):419–423. DOI:10.3760/cma.j.issn.0254−6450.2011.04.021.Zhang M, Wang XJ, Zhao ZT. Current epidemic status and issues on prevention and control of scrub typhus[J]. Chin J Epidemiol, 2011, 32(4): 419–423. DOI: 10.3760/cma.j.issn.0254−6450.2011.04.021. [9] 耿美玲, 操敏, 张锦海, 等. 建立染料法荧光定量PCR检测恙虫病东方体[J]. 中华卫生杀虫药械,2014,20(1):45–49. DOI:10.19821/j.1671−2781.2014.01.015.Geng ML, Cao M, Zhang JH, et al. Detection of Orientia tsutsugamushi by dye quantitative real-time PCR assay[J]. Chin J Hyg Insect Equip, 2014, 20(1): 45–49. DOI: 10.19821/j.1671−2781.2014.01.015. [10] 高玉峰, 孔文, 罗佳, 等. 恙虫病东方体与莫氏立克次体双重实时荧光PCR检测方法建立及其在鼠类检测中的应用[J]. 口岸卫生控制,2019,24(1):55–60. DOI:10.3969/j.issn.1008−5777.2019.01.019.Gao YF, Kong W, Luo J, et al. Establishment of a dual real-time fluorescent PCR detection method for detection of Orientia tsutsugamushi and Rickettsia Mooseriand its application in the detection of rodents[J]. Port Health Control, 2019, 24(1): 55–60. DOI: 10.3969/j.issn.1008−5777.2019.01.019. [11] Cordier C, Tattevin P, Leyer C, et al. Rickettsia sibirica mongolitimonae infection, Sri Lanka[J]. J Infect Dev Ctries, 2017, 11(8): 668–671. DOI: 10.3855/jidc.8743. [12] Kumar S, Kumar V, Chowdhary PK, et al. Rare and unusual presentation as immune thrombocytopenic Purpura in scrub typhus complicated by meningitis and acute kidney injury[J]. Indian J Crit Care Med, 2022, 26(6): 748–751. DOI: 10.5005/jp−journals−10071−24256. [13] Young PC, Hae CC, Lee KH, et al. Tsutsugamushi infection-associated acute rhabdomyolysis and acute renal failure[J]. Korean J Intern Med, 2003, 18(4): 248–250. DOI: 10.3904/kjim.2003.18.4.248. [14] 姚立农, 陆群英, 阮卫, 等. 五例恙虫病东方体感染基因型及其基因变异[J]. 中华实验和临床感染病杂志(电子版),2020,14(2):110–116. DOI:10.3877/cma.j.issn.1674−1358.2020.02.005.Yao LN, Lu QY, Ruan W, et al. Gene genotypes and variation of 5 cases with Orientia tsutsugamushi infection[J]. Chin J Exp Clin Infect Dis (Electr Ed), 2020, 14(2): 110–116. DOI: 10.3877/cma.j.issn.1674−1358.2020.02.005. [15] Chang WH, Kang JS, Lee WK, et al. Serological classification by monoclonal antibodies of Rickettsia tsutsugamushi isolated in Korea[J]. J Clin Microbiol, 1990, 28(4): 685–688. DOI: 10.1128/jcm.28.4.685−688.1990. -
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