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通过聚合酶链式反应(PCR)扩增后,作为检测微生物DNA生物传感器的空间稳定阳离子乳胶的桥连絮凝作用。

Bridging Flocculation of a Sterically Stabilized Cationic Latex as a Biosensor for the Detection of Microbial DNA after Amplification via PCR.

作者信息

Trinh Elisabeth, Batt Lauren J, Yue Qi, Du Ruiling, Jones Samuel T, Fielding Lee A

机构信息

Department of Materials, School of Natural Sciences, The University of Manchester, Oxford Road, Manchester, M13 9PL, United Kingdom.

Henry Royce Institute, The University of Manchester, Oxford Road, Manchester, M13 9PL, United Kingdom.

出版信息

Biomacromolecules. 2024 Mar 11;25(3):1629-1636. doi: 10.1021/acs.biomac.3c01187. Epub 2024 Feb 15.


DOI:10.1021/acs.biomac.3c01187
PMID:38361251
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10934273/
Abstract

There is a high demand for rapid, sensitive, and accurate detection methods for pathogens. This paper demonstrates a method of detecting the presence of amplified DNA from a range of pathogens associated with serious infections including Gram-negative bacteria, Gram-positive bacteria, and viruses. DNA is amplified using a polymerase chain reaction (PCR) and consequently detected using a sterically stabilized, cationic polymer latex. The DNA induces flocculation of this cationic latex, which consequently leads to rapid sedimentation and a visible change from a milky-white dispersion to one with a transparent supernatant, presenting a clear visible change, indicating the presence of amplified DNA. Specifically, a number of different pathogens were amplified using conventional or qPCR, including , , and Herpes Simplex Virus (HSV-2). This method was demonstrated to detect the presence of bacteria in suspension concentrations greater than 380 CFU mL and diagnose the presence of specific genomes through primer selection, as exemplified using methicillin resistant and methicillin susceptible . The versatility of this methodology was further demonstrated by showing that false positive results do not occur when a PCR of fungal DNA from is conducted using bacterial universal primers.

摘要

对病原体的快速、灵敏且准确的检测方法有很高的需求。本文展示了一种检测一系列与严重感染相关病原体的扩增DNA存在的方法,这些病原体包括革兰氏阴性菌、革兰氏阳性菌和病毒。使用聚合酶链反应(PCR)扩增DNA,随后使用空间稳定的阳离子聚合物乳胶进行检测。DNA诱导这种阳离子乳胶絮凝,进而导致快速沉淀,并出现从乳白色分散液到上清液透明的明显可见变化,呈现出清晰的可见变化,表明存在扩增的DNA。具体而言,使用常规PCR或定量PCR扩增了多种不同的病原体,包括……以及单纯疱疹病毒2型(HSV - 2)。该方法被证明能够检测悬浮液中浓度大于380 CFU/mL的细菌的存在,并通过引物选择诊断特定基因组的存在,以耐甲氧西林和对甲氧西林敏感的……为例进行了说明。通过使用细菌通用引物对来自……的真菌DNA进行PCR时未出现假阳性结果,进一步证明了该方法的通用性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0206/10934273/0ac128567baa/bm3c01187_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0206/10934273/78f872eb4c2c/bm3c01187_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0206/10934273/a1189ea5c252/bm3c01187_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0206/10934273/3b92922bc70e/bm3c01187_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0206/10934273/45fa7256ec99/bm3c01187_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0206/10934273/0ac128567baa/bm3c01187_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0206/10934273/78f872eb4c2c/bm3c01187_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0206/10934273/a1189ea5c252/bm3c01187_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0206/10934273/3b92922bc70e/bm3c01187_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0206/10934273/45fa7256ec99/bm3c01187_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0206/10934273/0ac128567baa/bm3c01187_0004.jpg

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[1]
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引用本文的文献

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Flexible Electrochemical Biosensor Using Nanostructure-Modified Polymer Electrode for Detection of Viral Nucleic Acids.

Biosensors (Basel). 2024-12-4

本文引用的文献

[1]
A Systematic Review of Second-Line Treatments in Antiviral Resistant Strains of HSV-1, HSV-2, and VZV.

Cureus. 2023-3-9

[2]
Visible label-free detection of bacterial DNA using flocculation of sterically stabilised cationic latexes.

J Mater Chem B. 2023-5-3

[3]
Broad-Spectrum Extracellular Antiviral Properties of Cucurbit[]urils.

ACS Infect Dis. 2022-10-14

[4]
Same-day confirmation of infection and antimicrobial susceptibility profiling using flow cytometry.

EBioMedicine. 2022-8

[5]
Physical Adsorption of Graphene Oxide onto Polymer Latexes and Characterization of the Resulting Nanocomposite Particles.

Langmuir. 2022-7-12

[6]
Diagnostic accuracy of rapid one-step PCR assays for detection of herpes simplex virus-1 and -2 in cerebrospinal fluid: a systematic review and meta-analysis.

Clin Microbiol Infect. 2022-12

[7]
How to: ECOFFs-the why, the how, and the don'ts of EUCAST epidemiological cutoff values.

Clin Microbiol Infect. 2022-7

[8]
Modular DNA Circuits for Point-of-Care Colorimetric Assay of Infectious Pathogens.

Anal Chem. 2021-10-19

[9]
Ultrasensitive and Highly Specific Lateral Flow Assays for Point-of-Care Diagnosis.

ACS Nano. 2021-3-23

[10]
Modern Tools for Rapid Diagnostics of Antimicrobial Resistance.

Front Cell Infect Microbiol. 2020-7-15

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