细胞因子IL-17作为牛结核病诊断标志物的研究及实时荧光定量PCR检测方法的建立

doi:10.16431/j.cnki.1671-7236.2018.06.003

Abstract

Abstract:

This study was aimed to explore the correlation between the transcription levels of cytokines (IL-6 and IL-17) and the infection of Mycobacterium bovis,and evaluate potential of these cytokines for bovine tuberculosis (bTB) diagnosis.Clinically TB-infected cattle and healthy cattle were screened by the skin tests and IFN-γ release assays.Heparinized blood from each animal was collected and used for isolation of the peripheral blood lymphocytes (PBMC).PBMCs were respectively incubated with bovine tuberculin (PPD-B),avian tuberculin (PPD-A),recombinant protein CFP-10-ESAT-6 (CE),PET or PBS at 37℃ for 6 h,and then the transcription levels of IL-6,IL-17 and IFN-γ mRNA were detected by Real-time PCR.As with PBS,PET could not stimulate the increase of cytokine mRNA level,which indicated the influence of PET contained in CE was negligible.PPD-B,PPD-A or CE significantly increased mRNA levels of IL-17 and IFN-γ in PMBCs from TB-infected cattle (P<0.05), of which PPD-B showed the highest stimulus intensity,while CE exhibited the best stimulus specificity.Under the condition that CE was selected as optimal stimulus,the correlation between IL-17 and respective IFN-γ mRNA levels were observed (spearman r=0.79),and then Real-time PCR assays of IL-17 and IFN-γ were preliminary developed.The detection rate of IL-17-based method on 14 clinical bTB-positive samples was 85.7%,which was higher than 71.4% of IFN-γ.Taken together,these findings supported the IL-17 mRNA level in PBMC was related with bTB infection,and demonstrated that measurement of IL-17 mRNA by Real-time PCR was potential for bTB diagnosis.

Key words: bovine tuberculosis; diagnosis; IL-17; IFN-γ; Real-time PCR

CLC Number:

S855.2

GAO Xintao, JIA Hong, HOU Shaohua, GUO Xiaoyu, YUAN Weifeng, JIANG Yitong, ZHU Hongfei, XIN Ting. Research on the Potential of IL-17 as a Biomarker of Bovine Tuberculosis and Establishment of Its Real-time PCR Assay[J]. , 2018, 45(6): 1447-1453.

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Research on the Potential of IL-17 as a Biomarker of Bovine Tuberculosis and Establishment of Its Real-time PCR Assay

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