热带病与寄生虫学

热带病与寄生虫学 ›› 2021, Vol. 19 ›› Issue (1): 15-18.

• 论著 • 上一篇    下一篇

恙虫病东方体目视LAMP现场化检测方法的建立

陶志勇,UMAR Ibrahim,吕樵岚,刘韦萍,张培艺,靳小霞,方强,夏惠   

  1. 蚌埠医学院病原生物学教研室,安徽蚌埠233030
  • 出版日期:2021-02-20 发布日期:2021-02-04
  • 通信作者: 夏惠,E-mail:Xiahui912@163.com
  • 作者简介:陶志勇,男,博士,副教授,研究方向:病原体的实验诊断。E-mail:taozhiyong@bbmc.edu.cn
  • 基金资助:
    安徽省高等学校科研创新平台团队项目(2016-40);安徽省高等学校自然科学研究重点项目(KJ2017A222);国家级大学生创新项目(201810367026);安徽省高等学校省级质量工程基层教研室示范项目(2018jyssf097);蚌埠市-蚌埠医学院联合科技攻关项目(BYLK201828)

Establishment of a field visual approach for detecting Orientia tsutsugamushi in chigger mites by LAMP device#br#

TAO Zhi-yong,UMAR Ibrahim,LÜ Qiao-lan, LIU Wei-ping ,ZHANG Pei-yi ,JIN Xiao-xia, FANG Qiang ,XIA Hui   

  1. Department of Microbiology and Parasitology Bengbu Medical College,Bengbu233030, Anhui Province ,China
  • Online:2021-02-20 Published:2021-02-04

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摘要： 目的 建立基于“染料-蜡丸”的目视LAMP检测恙螨中恙虫病东方体的方法,为恙虫病的防治提供新工具。方法 选择Ot GroEL 基因作为检测靶序列,在线设计并合成LAMP引物。构建rOt-GroEL 重组质粒作为阳性对照。建立基于SYBR Green I的“染料-蜡丸”目视LAMP,并联合便携式LAMP仪使其利于现场应用。结果 新设计的引物针对Ot GroEL 基因的237 bp靶区域,所构建的重组质粒rOt-GroEL 测序结果正确。在便携式LAMP仪上运用“染料-蜡丸”目视LAMP法,其检测下限为1×102 拷贝质粒。结论 成功建立了一种可与便携式LAMP仪联合使用的检测恙螨体内恙虫病东方体的“染料-蜡丸”目视LAMP法。

关键词: 恙虫病, 恙虫病东方体, 恙螨, 检测方法, 便携式LAMP仪

Abstract: Objective To establish a field visual method for detecting the Orientia tsutsugamushi (Ot) in chigger mites using self-prepared dye-wax capsule kit on portable loop-mediated isothermal amplification (LAMP) device basis for developing a new field tool in control and prevention of scrub typhus.Methods The GroEL gene was selected as the target sequence,and the primer for LAMP detection was designed online.A recombinant plasmid rOt- GroEL was constructed as the positive control.SYBR Green I based dye-wax capsule kit was used to establish a close-tube visual LAMP targeting GroEL.Field visual detection was simulated and verified using portable LAMP device. Results A set of LAMP primers specific to the Ot GroEL gene was chosen to amplify the 237 bp target region. Sequencing results showed that the recombinant rOt-GroEL plasmid was successfully constructed.Detection limit of the plasmid was 1×102 copies using dye-wax capsule kit combined with portable LAMP device.Conclusion A close-tube visual LAMP method targeting Ot GroEL was established,which could be used for field visual detecting Ot in chigger mites with portable LAMP device.

Key words: Scrub typhus , Orientia tsutsugamushi , Chigger mite , Detection method , Portable LAMP device

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