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

doi:10.3969/j.issn.1674-5817.2018.05.005

Abstract

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)

CLC Number:

R-332Q95-33

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.

References

[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.