Wang Yu, Zhao Xue, Cheng Jinzhi, Tang Xiaomin, Chen Xu, Yu Honglan, Li Shijun
Department of Clinical Laboratory, The First People's Hospital of Guiyang, Guiyang, China.
Laboratory of Bacterial Infectious Disease of Experimental Center, Guizhou Provincial Centre for Disease Control and Prevention, Guiyang, China.
Front Microbiol. 2021 Jun 10;12:681488. doi: 10.3389/fmicb.2021.681488. eCollection 2021.
is an increasingly opportunistic pathogen that causes serious invasive candidiasis threatening a patient's life. Traditional methods to detect infection depends on time-consuming, culture-based gold-standard methods. So, we sought to establish a new method that could detect target pathogens quickly, accurately, and straightforwardly. Herein, a combination of multiple cross displacement amplification (MCDA) and lateral flow biosensors (LFB) was employed to detect In the MCDA system, 10 primers were designed to identify the specific genes of and amplify the genes in an isothermal amplification device. Then, MCDA amplification reaction products could be identified visibly by color change, and all the amplification products would be tested by LFB with no special equipment. The results demonstrated that the optimal reaction condition of -MCDA assay was 64°C within 30 min, and only 10 fg DNA was required in each reaction. No cross-reaction was found between strains and non- strains. For 300 sputum samples, the results showed that MCDA-LFB assay could rapidly and successfully detect all of the -positive (28/300) samples detected by the gold-standard method. The entire procedure, including specimen processing (40 min), isothermal reaction (30 min) and result reporting (within 2 min), could be completed within 75 min. Briefly, the study results demonstrated that the detection ability of MCDA-LFB assay was better than culture methods with more simplicity, rapidity, sensitivity and specificity. Hence, MCDA-LFB strategy is an effective tool to rapidly detect in clinical samples, especially in resource-poor areas.
是一种越来越具机会致病性的病原体,可引起严重的侵袭性念珠菌病,威胁患者生命。传统的检测感染方法依赖于耗时的基于培养的金标准方法。因此,我们试图建立一种能够快速、准确且直接地检测目标病原体的新方法。在此,采用多重交叉置换扩增(MCDA)和侧向流动生物传感器(LFB)相结合的方法来检测。在MCDA系统中,设计了10条引物来识别的特定基因,并在等温扩增装置中扩增这些基因。然后,MCDA扩增反应产物可通过颜色变化肉眼识别,所有扩增产物无需特殊设备即可通过LFB进行检测。结果表明,-MCDA检测的最佳反应条件是在64°C下30分钟,每个反应仅需10 fg DNA。菌株与非菌株之间未发现交叉反应。对于300份痰液样本,结果显示MCDA-LFB检测能够快速且成功地检测出所有通过金标准方法检测为阳性(28/300)的样本。整个过程,包括样本处理(40分钟)、等温反应(30分钟)和结果报告(2分钟内),可在75分钟内完成。简而言之,研究结果表明MCDA-LFB检测的检测能力优于培养方法,具有更高的简便性、快速性、敏感性和特异性。因此,MCDA-LFB策略是一种快速检测临床样本中尤其是资源匮乏地区的有效工具。