Pathogen Detection Of AMR Genes
Delannoy, S. et al. "High throughput screening of antimicrobial resistance genes in Gram-negative seafood bacteria." Microorganisms (2022)
Objective: investigate antimicrobial resistance genes (ARGs) and mobile genetic elements (MGEs) in seafood bacteria in or to estimate antimicrobial resistance (AMR) burden conveyed by the marine sectorUtilization of Standard BioTools™ products: • Biomark™ HD system
• 96.96 Dynamic Array™ IFCs
• Real-Time PCR Analysis SoftwareConclusions: • Only 4.5% of isolates displayed ARG-MGE-associated patterns, which does not support the hypothesis of high-level human exposure through either direct exposure or seafood consumption
TALK WITH US
SHRIMP CASE STUDY
Objective: The objective of this study was to measure gene expression levels of Hemocyte immune response genes when exposed to White Spot Syndrome Virus (WSSV) and Fusarium solani (fungal) in order to understand the molecular mechanisms driving shrimp-pathogen interactions to develop strategies to control disease outbreaks in shrimp production systems with L. vannamei. Utilization of Standard BioTools products: Biomark SystemNanoFlex 4-IFC Controller96.96 dynamic arraysReal-Time PCR Analysis Software Conclusions: Genes that were monitored showed up regulation and down regulation over the time course study. Results reported can serve as immunomarkers for mortality events and could help reduce the loss in shrimp production systems
Results
• Eight ARG markers detected positive, including sulfonamide resistance (sul1) in five of 50 Pseudomonas isolates, all in shrimp
• Six MEG and nine IS targets detected positive
• 76 of 268 strains (28.4%) gave positive signal for either ARG or MGE
• 12 of 268 (4.5%) gave positive signal for both ARG and MGE
Conclusions
• ARB and ARGs decrease as the distance from sources of contamination increases
• Standard BioTools high-throughput qPCR is a fast and convenient method for simultaneous investigation of large numbers of genes and hundreds of samples
• The technology is easy to implement and easily adaptable and new relevant markers can be added to accommodate an emergence of new ARGs
• Powerful tool for ARG and MGE investigation in complex DNA samples
Results and conclusions
Methods
• Bacterial strains were isolated from skin and gills of cod, flat fishes, horse mackerel, shellfish and shrimp
• 74 ARGs and MGEs identified as best for this study
• TaqMan™ chemistry used on 96.96 IFC leveraging FAM™ and HEX™
• Positive controls used to validate the qPCR results consisted of already positively identified strains of isolated DNA by whole genome sequencing (WGS) or artificially made plasmid constructs
Background
• AMR is a global public health safety issue because resistance can reduce treatment options in bacterial infections
• These researchers wanted to analyze the prevalence of AMR and antimicrobial-resistant bacteria (ARB) in several different aquatic species
• Researchers isolated Gram-negative bacteria from five different aquatic species and analyzed them for the presence of ARGs