PulseNet: Molecular Subtyping of Foodborne Pathogens

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What is the significance of typing in epidemiologic (infectious disease) investigations, particularly in linking food source isolate to clinical isolate?

Typing helps to link food source isolate to clinical isolate, such as linking E. coli O157:H7 strain found in human faeces to that found in hamburger patties.

How does PulseNet contribute to the detection of foodborne outbreaks?

PulseNet detects outbreaks through surveillance, linking apparently sporadic cases to identify an outbreak, and identifies related cases, separating them from unrelated ones.

What is the role of molecular subtyping techniques in outbreak investigations?

Molecular subtyping techniques assist in rapidly identifying the source of outbreaks, stopping the sale of contaminated food while the outbreak is still small.

How does PulseNet USA coordinate with other agencies to investigate foodborne outbreaks?

PulseNet USA coordinates with state health departments, local health departments, and federal agencies (CDC, USDA/FSIS, FDA) to investigate foodborne outbreaks.

What is the significance of phenotyping methods in molecular epidemiology?

Phenotyping methods, such as serotyping and biotyping, provide essential information for identifying and characterizing pathogens, facilitating the investigation of outbreaks.

How does molecular epidemiology aid in the identification of foodborne pathogens?

Molecular epidemiology aids in the identification of foodborne pathogens through the use of molecular subtyping techniques, such as PCR and DNA fingerprinting.

What is the primary goal of PulseNet participants in terms of molecular subtyping of foodborne disease-causing bacteria?

To distinguish strains of bacteria at the DNA level.

What is the purpose of identifying matching subtypes of pathogens in different patients?

To identify the start of an outbreak and initiate an investigation.

What is the concept of 'typeability' in molecular subtyping, and what does it entail?

Typeability refers to the ability of a typing method to type all organisms, meaning all isolates can be distinguished.

What is the role of antibodies in serotyping, and which bacterial structures do they target?

Antibodies are used to target specific structures such as O (lipopolysaccharide), K (capsule), and H (flagella).

What is phage typing, and what is required for this method?

Phage typing is a phenotyping method that uses bacterial viruses (phages) to identify specific bacterial strains, requiring phage stocks.

What is the significance of antibiotic resistance profiles in phenotyping, and what do they indicate?

Antibiotic resistance profiles indicate the susceptibility of bacteria to different antibiotics, aiding in strain characterization.

What is the primary purpose of typing bacteria in the context of food microbiology?

To track the source and spread of food poisoning outbreaks and to track the source of spoilage microbes in food processing environments.

What is the difference between phenotyping and genotyping in molecular food microbiology?

Phenotyping involves identifying a microbe based on its physical and biochemical characteristics, while genotyping involves identifying a microbe based on its genetic makeup.

What is the significance of Sanger sequencing in the context of DNA sequencing?

Sanger sequencing is a traditional DNA sequencing technology that has been largely replaced by newer, high-throughput sequencing technologies known as Next-Generation Sequencing (NGS) technologies.

What is metagenomics analysis, and how is it related to genome sequencing?

Metagenomics analysis is a culture-independent approach that involves sequencing the genomes of microbial communities from environmental samples, such as food or soil.

How have changes in food consumption and practices in the United States affected the investigation of food poisoning outbreaks?

The shift from traditional point source outbreaks to more diffuse, widespread outbreaks has made it more challenging to detect and track foodborne illness.

What is the significance of PulseNet in the context of foodborne pathogen identification?

PulseNet is a network of laboratories that use standardized molecular subtyping techniques to identify and track foodborne pathogens.

Study Notes

PulseNet and Typing Methods

  • PulseNet was established in 1996 and performs standardized molecular subtyping of foodborne disease-causing bacteria.
  • The subtyping involves distinguishing strains of bacteria, such as Escherichia coli O157:H7, Salmonella, Shigella, Listeria, Campylobacter, Vibrio, and Cronobacter, at the DNA level.
  • The goal is to identify the start of an outbreak by looking for matching subtypes of pathogens in different patients.

Typing Methods Evaluation

  • Two key factors to consider when evaluating typing methods are typeability and discriminatory power.
  • Typeability refers to whether all organisms can be typed using a particular method.
  • Discriminatory power refers to a method's ability to distinguish unrelated bacterial isolates but not clones.

Phenotyping Methods

  • Phenotype is the result of the genotype and environmental influences, and it refers to any observable characteristic or trait.
  • Serotyping uses antibodies to identify O (lipopolysaccharide), K (capsule), and H (flagella) antigens.
  • There are 2500 serotypes of Salmonella, and E. coli O157:H7 is a specific serotype.

Applications of Typing

  • Typing helps in epidemiologic investigations by linking food source isolates to clinical isolates.
  • It detects outbreaks through surveillance and links apparently sporadic cases to identify an outbreak.
  • Typing identifies related cases and separates them from unrelated ones, assisting in rapidly identifying the source of outbreaks.
  • It helps stop the sale of contaminated food causing an outbreak while it is still small.

PulseNet USA

  • PulseNet is a national network of public health and food regulatory agency laboratories coordinated by the Centers for Disease Control and Prevention (CDC).
  • The network consists of state health departments, local health departments, and federal agencies (CDC, USDA/FSIS, FDA).

Molecular Food Microbiology Overview

  • Molecular food microbiology involves typing (subtyping), DNA sequencing, and genome sequencing.
  • Typing methods include phenotyping and genotyping.
  • DNA sequencing technologies include Sanger and new sequencing technologies (NGS).

Why Type Bacteria?

  • Typing bacteria is necessary to track the source and spread of food poisoning outbreaks.
  • It is also important for surveillance to track the source of spoilage microbes in food processing environments.

PulseNet is a network that uses molecular subtyping to identify and track foodborne disease-causing bacteria. It helps identify outbreaks by matching subtypes of pathogens in different patients. This leads to investigations and control of foodborne outbreaks.

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