弓形虫生物检测靶标miR-191的鉴定

doi:10.3969/j.issn.1674-5817.2018.05.005

实验动物与比较医学 ›› 2018, Vol. 38 ›› Issue (5): 350-355.DOI: 10.3969/j.issn.1674-5817.2018.05.005

弓形虫生物检测靶标miR-191的鉴定

温福利¹, 郑和平¹, 党源¹, 薛来恩¹, 赵东岳^1,2

1.南京军区福州总医院比较医学科,福州 350025;
2.福建师范大学生命科学学院,福州 350001

收稿日期:2018-05-31 出版日期:2018-10-25 发布日期:2018-10-25
作者简介:温福利(1987-),男,技师,从事实验动物寄生虫研究。E-mail:wen_fuli@163.com
基金资助:
福州总医院临床应用研究专项(2015L01)和福建省教育厅A类项目(JAT170137)

Identification of the Target miR-191 for the Biological Detection of Toxoplasma Gondii

WEN Fu-li¹, ZHENG He-ping¹, Dang Yuan¹, Xue Lai-en¹, ZHAO Dong-yue²

1. Department of Comparative Medicine,Fuzhou General Hospital of Nanjing Command,PLA,Fuzhou 350025,China;
2. College of Life Sciences,Fujian Normal University,Fuzhou 350001,China

Received:2018-05-31 Online:2018-10-25 Published:2018-10-25

摘要/Abstract

摘要： 目的建立弓形虫miRNA探针定量检测法。方法采用高通量测序技术筛选出不同弓形虫虫株间表达量高且差异不明显潜在靶标,并对其不同时间点(0 d、1 d、3 d、5 d、7 d)外周血的表达及诊断效能进行研究。结果高通量测序表明,miR-252、miR-3641、miR-509-5p、miR-1285、miR-140、miR128-1、miR-191、miR-3426具有作为血液中弓形虫生物检测靶标的潜在性。通过对感染弓形虫RH株和M49株小鼠的miR-191诊断效能分析后表明,敏感性分别为85%和95%,特异性均为100%。根据miR-191制备纳米金探针,对不同时间点感染弓形虫RH株和ME49株大鼠的血液靶标miR-191进行定量分析表明,感染后7 d内感染弓形虫miR-191表达情况相对稳定。制作弓形虫RH株、弓形虫ME49株、伯氏疟原虫、约氏疟原虫、隐孢子虫,鼠肝炎病毒和金黄色葡萄球菌感染的小鼠模型,与对照组比较分析表明,miR-191在感染弓形虫 RH株和ME49 株的小鼠模型中出现高表达,而在感染细菌、病毒和其它寄生虫的小鼠模型中均未出现表达。结论弓形虫miR-191具有作为检测靶标的应用价值。

关键词: 弓形虫, 高通量测序, miRNA

Abstract: Objective To establish the quantitative detection method of microRNAs (miRNAs) probe of Toxoplasma gondii. Methods High-throughput sequencing technology was used to screen out the high-level and non-significant potential targets between different strains of Toxoplasma gondii, and the expression and diagnostic efficiency of peripheral blood at different time points (0 d, 1 d, 3 d, 5 d, 7 d) were studied. Results High-throughput sequencing revealed that miR-252, miR-3641, miR-509-5p, miR-1285, miR-140, miR128-1, miR-191 and miR-3426 had potential as biological detection targets for Toxoplasma gondii. The miR-191 diagnostic efficiency of mice infected with Toxoplasma gondii RH strain and M49 strain showed that the specificity were 100%, and the sensitivity were 85% and 95%, respectively. Nanoscale gold probes were prepared by miR-191. The quantitative analysis of target miR-191 in blood of rats infected with Toxoplasma gondii RH strain and ME49 strain at different time points showed that miR-191 expression was relatively stable within 7 days after infection.The infection models of mice with Toxoplasma RH strain, Toxoplasma ME49 strain, Plasmodium bergi, Plasmodium yoelii, Cryptosporidium, mouse hepatitis virus and Staphylococcus aureus were made respectively. Compared with those of in control group, the miR-191 was highly expressed in mice infected with RH and ME49 strains of Toxoplasma gondii and no expression was found in mouse models infected with bacteria, virus and other parasites. Conclusions The miR-191 of Toxoplasma gondii has the application value as a detection target.

Key words: Toxoplasma gondii, Biological detection, microRNA (miRNAs)

中图分类号:

R-332Q95-33

温福利, 郑和平, 党源, 薛来恩, 赵东岳. 弓形虫生物检测靶标miR-191的鉴定[J]. 实验动物与比较医学, 2018, 38(5): 350-355.

WEN Fu-li, ZHENG He-ping, Dang Yuan, Xue Lai-en, ZHAO Dong-yue. Identification of the Target miR-191 for the Biological Detection of Toxoplasma Gondii[J]. Laboratory Animal and Comparative Medicine, 2018, 38(5): 350-355.

参考文献

[1] Xiao J,Li Y,Prandovszky E,et al.MicroRNA-132 dysregulation in Toxoplasma gondii infection has implications for dopamine signaling pathway[J].Neuroscience,2014,268(21):128-138.
[2] 吕允凤.弓形虫IgM抗体检测试剂临床研究要点[J].中国药物警戒,2014,15(10):632-634.
[3] Lao K,Xu NL,Yeung V,et al.Multiplexing RT-PCR for the detection of multiple miRNAs species in small samples[J].Biochem Bioph Res Commun,2006,343(1):85-89.
[4] Li YE,Kannan G,Pletnikov MV,et al.Chronic infection of Toxoplasma gondii downregulates miR-132 expression in multiple brain regions in a sex-dependent manner[J].Parasitology,2015,142(4):623-632.
[5] Thirugnanam S,Rout N,Gnanasekar M.Possible role of Toxoplasma gondii in brain cancer through modulation of host microRNAs[J].Infect Agents Cancer,2013,8(1):1-8.
[6] 谷俊朝,甘绍伯.弓形虫病的现状和展望[J].北京医学,2008,30(12):729-731.
[7] Chen X,Ba Y,Ma L,et al.Characterization of miRNAs in serum:a novel class of biomarkers for diagnosis of cancer and other diseases[J].Cell Res,2008,18(10):997-1006.
[8] Mitchell PS,Parkin RK,Kroh EM,et al.Circulating miRNAs as stable blood-based markers for cancer detection[J].Proc Natl Acad Sci U S A,2008,105(30):10513-10518.
[9] Bommer W.The life cycle of virulent toxoplasma in cell cultures[J].Aust J Exp Biol Med Sci,1969,47(4):505-512.
[10] Zeiner GM,Norman KL,Thomson JM.Toxoplasma gondii infection specifically increases the levels of key host microRNAs[J].PLoS One,2010,5(1):8742.
[11] Wang J,Liu X,Jia B,et al.A comparative study of small RNAs in Toxoplasma gondii of distinct genotypes[J].Parasite Vector,2012,5(1):186.
[12] Jia B,Chang Z,Wei X,et al.Plasma microRNAs are promising novel biomarkers for the early detection of Toxoplasma gondii infection[J].Parasite Vector,2014,7(1):433.
[13] 何文蕾,杨文杰,李媛媛,等.基于银染增强的纳米金探针定量检测microRNA[J].生物化学与生物物理进展,2008,35(11):1332-1338.

弓形虫生物检测靶标miR-191的鉴定

Identification of the Target miR-191 for the Biological Detection of Toxoplasma Gondii

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