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      Sputum miR-155 as a biomarker in detection with Mycobacterium tuberculosis infection

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          Abstract

          Objective To analyze Mycobacterium tuberculosis ( M. tb) strain H37Rv induced differential expression miRNAs from host macrophage, and we explore the diagnosis value of M.tb infection as a biomarker in clinic.

          Methods The Mycobacter tuberculosis (M.tb) strain H37Rv was incubated and cultured with human macrophage line THP-1. The total RNA was extracted from the THP-1 cells in 12 h, 24 h and 48 h. The miRNA of groups was detected by miRNA microarray chips and the differential expressed miRNAs were analyzed for verification experiments furtherly. The rt-PCR was used to detect the differential expressed miRNAs from infected THP-1 by M.tb strains, isolated from 120 clinical sputum samples, respectively. Following, the RNA was extracted from sputum sample to detect the miRNA using rt-PCR and this detecting efficiency of several methods was compared.

          Results The data showed the expression level of six miRNA have a significant change, that miR-155, miR-29b-1*, miR-150, miR-146a, miR-212 and miR-483-5p. The six miRNAs were detected from THP-1 cells, were infected by 120 clinical M. tb strains from different areas for 24 h. The results showed miR-155 was a higher level than others in the tests. After that the miR-155 as a biomarker for M.tb infection diagnosis, the RNA was extracted from 68 sputum samples to detect the level of miR-155. The sensitivity and specificity of miR-155 detectionwere 94.1% and 93.8%, respectively. The sensitivity was higher than that of sputum smear in 22.1% and tuberculosis antibody in 83.8% significantly ( P<0.05). But there was no significant difference with ELISPOT in 97.1% ( P>0.05). The ranking of sputum smear was positive correlation with miR-155 detection in sputum ( R2=0.743, P<0.05).

          Conclusions The data suggested that miR-155 detection is a viral significance for active TB infection diagnosis and bacteria-loads evaluation indirectly as an efficiency biomarker.

          Abstract

          摘要: 目的 分析结核分枝杆菌 H37Rv 感染诱导宿主巨噬细胞表达 miRNA 的变化, 进而验证差异 miRNA 对结核 分枝杆菌感染的临床诊断意义。 方法 使用 H37Rv 菌株感染人巨噬细胞系 THP-1 细胞后, 收集 12 h、24 h 和 48 h 细胞 并提取总 RNA, 进行 miRNA 微阵列芯片分析; 再将临床分离的 120 株不同个体来源的结核分枝杆菌菌株与 THP-1 细胞 共培养, 使用实时定量 PCR (real time-PCR, rt-PCR)验证差异 miRNA 的表达水平; 随后收集结核患者的痰液标本处理 后, 提取总 RNA, 进行实时荧光定量 PCR 检测痰液中的差异表达的 miRNA 水平。 结果 H37Rv 体外刺激 THP-1 细胞, 使用 miRNA 微阵列芯片分析, 结果显示:miR-155、miR-29b-1*、miR-150、miR-146a、miR-212 和 miR-483-5p 共计 6 个 miRNAs 水平在 3 个时间点升高均较为明显; 将来自不同区域的 120 株结核菌株分别与 THP-1 细胞的共培养 24 h, 检测 发现 miR-155 表达水平最高; 以 miRNA-155 为检测靶标, 对 68 例结核患者的痰液进行检测, 灵敏度和特异度分别为 94.1% 和 93.8%。其灵敏度高于痰涂片的 22.1% 和结核抗体的 83.8% , 差异具有统计学意义 ( P<0.05) 。但与 ELISpot 法 的 97.1%, 差异无统计学意义 ( P>0.05) 。在对痰涂片阳性结果与 miRNA 检测结果相关性分析中, 发现 miR-155 其表达 水平与痰涂片阳性等级呈正相关 ( R2=0.743, P<0.05) 。 结论 此研究提示 miR-155 能够作为活动性结核诊断的有效靶 标, 且对间接评估患者肺部荷菌量有重要意义。

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          Author and article information

          Journal
          CTM
          China Tropical Medicine
          China Tropical Medicine (China )
          1009-9727
          1 April 2020
          1 May 2020
          : 20
          : 4
          : 359-363
          Affiliations
          1Department of Respiratory and Critical Care Medicine, Longgang District Center Hospital of Shenzhen, Shenzhen, Guangdong 518000, China
          2Shenzhen Center for Chronic Disease Control, Shenzhen, Guangdong 518000, China
          3Basic Medical Laboratory, the Second Affiliated Hospital of Wannan Medical College, Wuhu, Anhui 241000, China
          Author notes
          Corresponding author: TANG Xiaolei, E-mail: 278471655@ 123456qq.com
          Article
          j.cnki.46-1064/r.2020.04.14
          10.13604/j.cnki.46-1064/r.2020.04.14
          © 2020 Editorial Department of China Tropical Medicine

          This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 Unported License (CC BY-NC 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. See https://creativecommons.org/licenses/by-nc/4.0/.

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