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使用多重交叉置换扩增结合基于纳米颗粒的侧向流动技术进行快速检测

Rapid Detection of Using Multiple Cross Displacement Amplification Combined With Nanoparticles-Based Lateral Flow.

作者信息

Jiang Luxi, Li Xiaomeng, Gu Rumeng, Mu Deguang

机构信息

Department of Respiratory Medicine, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, China.

Graduate School of Clinical Medicine, Bengbu Medical College, Bengbu, China.

出版信息

Front Cell Infect Microbiol. 2021 Apr 13;11:622402. doi: 10.3389/fcimb.2021.622402. eCollection 2021.

Abstract

is an opportunistic, ubiquitous, saprophytic mold which can cause infection in the lungs, nose, eyes, brain, and bones in humans, especially in immunocompromised patients. However, it is difficult to diagnose infection quickly. Here, we introduce a new detection method, namely multiple cross displacement amplification (MCDA) combined with nanoparticle-based lateral flow biosensor (LFB) (MCDA-LFB), which was proved to be fast, reliable, and simple for detecting . We designed a set of 10 primers targeting the gene annexin ANXC4 of . The best MCDA condition is 66 °C for 35 min. The minimum concentration that can be detected by this method was 10 fg. In the case of 100 sputum samples, 20 (20%) and 15 (15%) samples were positive by MCDA-LFB and PCR method, respectively. MCDA-LFB and traditional culture method showed the same results. Compared with the culture method, the diagnostic accuracy of MCDA-LFB can reach 100%. It showed that the MCDA-LFB method has better detection ability than the PCR method. We found that the whole process could be controlled within 60 min including the preparation of DNA (20 min), MCDA reaction (35 min) and results reporting (2 min). These results show that this assay is suitable for the rapid, sensitive and specific detection of in clinical samples.

摘要

是一种机会性、无处不在的腐生霉菌,可导致人类肺部、鼻子、眼睛、大脑和骨骼感染,尤其是免疫功能低下的患者。然而,很难快速诊断感染。在此,我们介绍一种新的检测方法,即多重交叉置换扩增(MCDA)与基于纳米颗粒的侧向流动生物传感器(LFB)相结合(MCDA-LFB),该方法被证明对检测……快速、可靠且简单。我们设计了一组针对……膜联蛋白ANXC4基因的10条引物。最佳MCDA条件是66℃反应35分钟。该方法可检测的最低浓度为10 fg。在100份痰液样本中,MCDA-LFB和PCR方法分别有20份(20%)和15份(15%)样本呈阳性。MCDA-LFB与传统培养方法结果相同。与培养方法相比,MCDA-LFB的诊断准确率可达100%。结果表明,MCDA-LFB方法比PCR方法具有更好的检测能力。我们发现整个过程可在60分钟内完成,包括DNA制备(20分钟)、MCDA反应(35分钟)和结果报告(2分钟)。这些结果表明该检测方法适用于临床样本中……的快速、灵敏和特异性检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9765/8076636/2c762c51cb74/fcimb-11-622402-g001.jpg

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