纳米金探针对铜绿假单胞菌的比色检测。

IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal, genetic engineering & biotechnology Pub Date : 2023-06-27 DOI:10.1186/s43141-023-00527-4
Zahra Mousivand, Fatemeh Haddadi, Hossein Kamaladini
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引用次数: 1

摘要

背景:由于分子方法相对于生化方法的优势,利用分子方法诊断假单胞菌等院内感染,可为正确诊断和治疗感染,预防感染引起的进一步并发症提供一种合适、快速的方法。本文介绍了一种基于纳米颗粒的检测技术的发展,该技术可用于铜绿假单胞菌的敏感和特异性脱氧核糖核酸诊断。设计了16S rDNA高变区硫代寡核苷酸特异性探针,并应用于该细菌的比色检测。结果:金纳米探针核酸序列扩增结果表明,探针在目标脱氧核糖核酸存在的情况下附着在金纳米颗粒上。它导致金纳米颗粒以连接网络的形式聚集,导致颜色变化,并表明样品中存在目标分子,这可以通过肉眼观察到。此外,金纳米粒子的波长从524 nm变化到558 nm。利用铜绿假单胞菌的4个特异性基因(oprL、oprI、toxA和16S rDNA)进行多重聚合酶链反应。评估两种技术的敏感性和特异性。观察结果表明,两种技术的特异性均为100%,多重聚合酶链反应和比色法测定基因组脱氧核糖核酸的灵敏度分别为0.5 ng/μL和0.01 ng/μL。结论:比色法检测的灵敏度比16SrDNA基因聚合酶链反应高50倍左右。我们的研究结果证明是高度特异性的,可能用于铜绿假单胞菌的早期检测。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Colorimetric bacteria sensing of Pseudomonas aeruginosa using gold nanoparticle probes.

Background: Due to the advantages of molecular methods over biochemical methods, the use of molecular methods for diagnosing nosocomial infections such as Pseudomonas can be an appropriate and rapid way to choose the right diagnosis and treatment of infection and prevent further complications caused by the infection. The present article provides a description of the development of a nanoparticle-based detection technique for sensitive and specific deoxyribonucleic acid-based diagnostic of Pseudomonas aeruginosa. Specific thiolated oligonucleotide probes for one of the hypervariable regions of the 16S rDNA gene were designed and applied for colorimetric detection of the bacteria.

Results: The results of gold nanoprobe-nucleic sequence amplification indicated the probe attached to gold nanoparticles in the presence of the target deoxyribonucleic acid. It caused aggregation of gold nanoparticles in the form of connected networks resulting in color change and indicating the presence of the target molecule in the sample, which could be observed by the naked eye. In addition, the wavelength of gold nanoparticles changed from 524 to 558 nm. Multiplex polymerase chain reactions were performed using four specific genes of Pseudomonas aeruginosa (oprL, oprI, toxA, and 16S rDNA). The sensitivity and specificity of the two techniques were assessed. According to the observations, the specificity of both techniques was 100%, and the sensitivity was 0.5 ng/μL and 0.01 ng/μL of genomic deoxyribonucleic acid for multiplex polymerase chain reaction and colorimetric assay, respectively.

Conclusions: The sensitivity of colorimetric detection was about 50 times higher than the polymerase chain reaction using the 16SrDNA gene. The results of our study proved to be highly specific with potential use for early detection of Pseudomonas aeruginosa.

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