Advances in detection methods for viable Salmonella spp.: current applications and challenges

Zhuang, Linlin; Gong, Jiansen; Shen, Qiuping; Yang, Jianbo; Song, Chunlei; Liu, Qingxin; Zhao, Bin; Zhang, Yu; Zhu, Mengling

doi:10.1007/s44211-023-00384-8

Cited by 4 publications

(2 citation statements)

References 170 publications

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“…PMAxx Dye (20 mM) (Biotium, Inc., Hayward, CA, USA) was stored at −20 • C. Various concentrations of PMAxx (10,25,50,75, and 100 µM) were added to viable and dead S. Typhimurium at a concentration of 1.34 × 10 8 CFU/mL. After adding PMAxx, samples were incubated in the dark at room temperature for 10 min to allow penetration into damaged membranes.…”

Section: Verification Of the Pmaxx Treatmentmentioning

confidence: 99%

“…These chemicals effectively inhibit the detection of DNA from dead bacterial cells, thereby enhancing the accuracy of rapid detection methods. A recently improved version of PMA, called PMAxx, has shown enhanced efficacy in binding to DNA from dead cells and has been applied in various isothermal amplification methods and PCR to selectively detect viable Salmonella species [24,25]. However, research on PMAxx-RPA-based NALF for detecting viable Salmonella species has not been reported.…”

Section: Introductionmentioning

confidence: 99%

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Point-of-Care Diagnostic System for Viable Salmonella Species via Improved Propidium Monoazide and Recombinase Polymerase Amplification Based Nucleic Acid Lateral Flow

Lee,

2024

Diagnostics

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Salmonella species are prominent foodborne microbial pathogens transmitted through contaminated food or water and pose a significant threat to human health. Accurate and rapid point-of-care (POC) diagnosis is gaining attention in effectively preventing outbreaks of foodborne disease. However, the presence of dead bacteria can interfere with an accurate diagnosis, necessitating the development of methods for the rapid, simple, and efficient detection of viable bacteria only. Herein, we used an improved propidium monoazide (PMAxx) to develop a nucleic acid lateral flow (NALF) assay based on recombinase polymerase amplification (RPA) to differentiate viable Salmonella Typhimurium. We selected an RPA primer set targeting the invA gene and designed a probe for NALF. RPA-based NALF was optimized for temperature (30–43 °C), time (1–25 min), and endonuclease IV concentration (0.025–0.15 unit/µL). PMAxx successfully eliminated false-positive results from dead S. Typhimurium, enabling the accurate detection of viable S. Typhimurium with a detection limit of 1.11 × 102 CFU/mL in pure culture. The developed method was evaluated with spiked raw chicken breast and milk with analysis completed within 25 min at 39 °C. This study has potential as a tool for the POC diagnostics of viable foodborne pathogens with high specificity, sensitivity, rapidity, and cost-effectiveness.

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Section: Verification Of the Pmaxx Treatmentmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%