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 Staphylococcus aureus, Escherichia coli, and Herpes Simplex Virus (HSV-2). This method was demonstrated to detect the presence of bacteria in suspension concentrations greater than 380 CFU mL–1 and diagnose the presence of specific genomes through primer selection, as exemplified using methicillin resistant and methicillin susceptible Staphylococcus aureus. The versatility of this methodology was further demonstrated by showing that false positive results do not occur when a PCR of fungal DNA from C. albicans is conducted using bacterial universal primers.
| Original language | English |
|---|---|
| Pages (from-to) | 1629-1636 |
| Number of pages | 8 |
| Journal | Biomacromolecules |
| Volume | 25 |
| Issue number | 3 |
| Early online date | 16 Feb 2024 |
| DOIs | |
| Publication status | Published - 11 Mar 2024 |
Bibliographical note
AcknowledgmentsHSV-2 (Herpes Simplex Virus) was isolated, verified, and kindly donated by Dr. Carol Yates at the School of Medical Sciences, University of Manchester. The Medical Research Council and University of Manchester Doctoral Training Programme is thanked for the Ph.D. studentship for E.T. through Grants MR/N013751/1 and MR/R015767/1. The Biotechnology and Biological Sciences Research Council and the Manchester Doctoral Training Programme is thanked for the Ph.D. studentship for L.J.B. through Grant DTP3 2020-2025, Reference BB/T008725/1. The University of Manchester Electron Microscopy Centre is acknowledged for access to electron microscopy facilities. This work was supported by the Henry Royce Institute for Advanced Materials, funded through EPSRC Grants EP/R00661X/1, EP/S019367/1, EP/P025021/1, and EP/P025498/1.