Biomedical Informatics & Genomics Center, Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, China.
Research and Development Department, Qingdao Haier Biotech Co. Ltd, Qingdao, China.
Front Cell Infect Microbiol. 2021 Sep 22;11:721547. doi: 10.3389/fcimb.2021.721547. eCollection 2021.
is a major human pathogenic bacterium, resulting in a series of diseases, such as pneumonia, bacteremia, meningitis. However, it is hard to diagnose . quickly. In this study, the multiple cross displacement amplification (MCDA) and nanoparticle-based lateral flow biosensor (LFB) (MCDA-LFB) were combined to detect . , which has been proven to be reliable, rapid, and not complicated. On the basis of . outer membrane protein gene, 10 specific primers were designed. The best MCDA condition was 61°C for 1 h. The sensitivity of . -MCD-LFB assay showed, in the pure cultures, the minimum concentration of genomic DNA templates was 100 fg. The specificity of . -MCD-LFB assay showed only . templates were detected, and no cross-reactivity was found in non-. isolates and other species. In 56 sputum samples, with MCDA-LFB method and PCR detection, 21 samples were positive, which was in consistent with the traditional culture method. The accuracy of diagnosis of MCDA-LFB, in comparison with the traditional culture method and PCR detection, can reach 100%, indicating that the MCDA-LFB assay gains an advantage over the cultured-based method for target pathogen detection. In conclusion, the MCDA-LFB assay is suitable for the sensitive, rapid, and specific detection of . , which might be used as a potential diagnostic tool for . in basic and clinical laboratories.
是一种主要的人类致病菌,可导致一系列疾病,如肺炎、菌血症、脑膜炎。然而,它很难快速诊断。在这项研究中,将多重交叉置换扩增(MCDA)和基于纳米颗粒的侧流生物传感器(LFB)(MCDA-LFB)相结合,以检测,这已被证明是可靠、快速且不复杂的。基于外膜蛋白基因,设计了 10 个特异性引物。最佳的 MCDA 条件是 61°C 1 小时。检测的最低基因组 DNA 模板浓度为 100 fg。检测的特异性仅检测到模板,非和其他种属未发现交叉反应。在 56 份痰样本中,用 MCDA-LFB 方法和 PCR 检测,有 21 份为阳性,与传统培养方法一致。与传统培养方法和 PCR 检测相比,MCDA-LFB 方法的诊断准确性可达到 100%,表明 MCDA-LFB 检测方法优于基于培养的方法,适用于目标病原体的检测。总之,MCDA-LFB 检测法适用于的敏感、快速和特异性检测,可能成为基础和临床实验室检测的潜在诊断工具。