Salmonella PCR Assay Overview
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Questions and Answers

What is the primary purpose of the Thermo Scientific™ SureTect™ Salmonella species PCR Assay?

  • To detect and identify Salmonella species in food samples (correct)
  • To evaluate the nutritional content of food products
  • To assess the bacterial growth rate in food
  • To measure the temperature of food samples

What technology is utilized in the SureTect PCR assays for DNA amplification?

  • Real-time Polymerase Chain Reaction (PCR) (correct)
  • Sanger Sequencing
  • Gel Electrophoresis
  • Mass Spectrometry

Which item is NOT required as part of the SureTect System?

  • Homogenizer bag rack
  • DNA extraction kit (correct)
  • Homogenizer Laboratory Blender
  • Sterile water

Which component is not included in the PCR pellets of the SureTect assays?

<p>Fluorescent dyes for staining (C)</p> Signup and view all the answers

What temperature ranges are specified for the incubators used with the SureTect System?

<p>37°C ±1°C, 41.5°C ±1°C (C)</p> Signup and view all the answers

How does the Minor Groove Binder (MGB) affect the PCR probes in the SureTect assays?

<p>It enhances the probe-template DNA bond. (D)</p> Signup and view all the answers

What is the expected turnaround time for results when using the SureTect Salmonella species PCR Assay?

<p>1 hour and 20 minutes (D)</p> Signup and view all the answers

Which agar is recommended for the isolation and identification of Salmonella colonies?

<p>Oxoid Brilliance™ Salmonella Agar (A)</p> Signup and view all the answers

What is strongly recommended before implementing a test for an unvalidated sample type?

<p>Conduct a full evaluation of the enrichment procedure (A)</p> Signup and view all the answers

What compliance is advised when using PCR methods in the SureTect System?

<p>Compliance with Good Laboratory Practices (D)</p> Signup and view all the answers

Which broth has been validated for chocolate matrices during the AOAC-RI PTM validation?

<p>Oxoid Cocoa Samples Recovery (CSR) Broth (D)</p> Signup and view all the answers

What supplement is added to BPW for validating germination tests on sprouted seeds?

<p>20mg/l of novobiocin (C)</p> Signup and view all the answers

What is one of the required tools for sample processing mentioned in the content?

<p>Lysis Tube Rack (D)</p> Signup and view all the answers

Which instrument is compatible with SureTect Software for analysis of the SureTect assay?

<p>SureTect PikoReal Real-Time PCR Instrument (D)</p> Signup and view all the answers

What operating system is required for the Applied Biosystems 7500 Fast instrument's software?

<p>Windows 7 (B)</p> Signup and view all the answers

What should be done before adding a beef sample to the homogenizer bag?

<p>Add 1.5 litres of pre-warmed mTSB at 41.5°C. (A)</p> Signup and view all the answers

How should the enriched CSR Broth sample be treated prior to the lysis protocol for chocolate samples?

<p>It should be diluted 1:5 using sterile CSR Broth or Peptone Salt solution. (C)</p> Signup and view all the answers

What is the recommended incubation temperature for samples added to Buffered Peptone Water after homogenization?

<p>37°C ±1°C. (A)</p> Signup and view all the answers

Which of the following is a proper step for sampling areas treated with disinfectants?

<p>Pre-moisten with a neutralizing broth like Dey-Engley Broth. (D)</p> Signup and view all the answers

How long can samples be held at room temperature before proceeding to the next step?

<p>Up to 2 hours. (A)</p> Signup and view all the answers

Flashcards

SureTect Salmonella species PCR Assay

A real-time PCR test used to detect and identify Salmonella species in food and food manufacturing environmental samples.

Real-time PCR

A technique that amplifies DNA while simultaneously monitoring the reaction in real-time.

Probes

Short, DNA sequences used in PCR to detect specific target regions.

MGB

Minor Groove Binder, a molecule attached to probes that enhances the signal and reduces background noise.

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Solaris™ qPCR technology

The technology behind SureTect assays, using probes with MGB to improve PCR detection.

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AOAC-RI PTM Validation

A standardized process to assess the accuracy and reliability of a specific method for detecting Salmonella in different food matrices.

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Oxoid Cocoa Samples Recovery (CSR) Broth

A specific type of broth specifically validated for use with chocolate samples in the AOAC-RI PTM validation.

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BPW (ISO) supplemented with 20mg/l novobiocin

A specific growth medium validated for use with sprouted seeds in the AOAC-RI PTM validation. Novobiocin is an antibiotic that inhibits the growth of some bacteria.

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PikoReal Real-Time PCR Instrument

A machine used to carry out the SureTect Salmonella assay. It allows researchers to detect and quantify specific DNA sequences in real-time.

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Enrichment Broth

A liquid growth medium used to increase the number of bacteria in a sample, making them easier to detect.

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Homogenizer Bag

A specialized bag for blending and breaking down food samples to release bacteria for testing.

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Dilute Enriched Sample

To reduce the concentration of bacteria in a sample by adding more broth before analysis.

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Pre-moistened Swab

A sterile cotton swab soaked in a liquid to collect bacteria from surfaces.

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Buffered Peptone Water

A sterile liquid solution that is used to dilute and neutralize samples collected from surfaces.

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SureTect System Components

The SureTect System is a complete testing system for Salmonella detection. However, it doesn't include all necessary equipment. You will need to acquire additional items like a blender, homogenizer bag rack, incubator, and sterile water.

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Homogenizer

A laboratory device used to blend (homogenize) samples for analysis. It breaks down the sample structure to release Salmonella bacteria.

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Incubators

Specialized ovens used to control and maintain a specific temperature. They are essential for growing Salmonella cultures.

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Confirmation of Positive Results

After running the SureTect assay, it's important to confirm positive results by culturing the sample on a suitable medium like Oxoid Brilliance™ Salmonella Agar. This step helps isolate and identify Salmonella colonies.

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ISO Standards for Sample Preparation

Specific guidelines and procedures exist for preparing samples for Salmonella analysis, based on international standards like ISO 6887 and ISO 65794. These standards ensure consistent and reliable results.

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Study Notes

Thermo Scientific™ SureTect™ Salmonella species PCR Assay PT0100A

  • Intended for use with Thermo Scientific™ Piko Real™ Real-Time PCR Instrument or Applied Biosystems™ 7500 Fast Real-Time PCR System
  • Detects Salmonella species in food and food manufacturing environmental samples
  • Real-time Polymerase Chain Reaction (PCR) technology amplifies DNA for detection and analysis
  • Uses Solaris™ qPCR technology with Minor Groove Binder (MGB) probes for enhanced signal-to-noise ratio
  • Achieves results in approximately 1 hour and 20 minutes
  • Results displayed as positive or negative symbols with amplification plots
  • Assay includes reagents for bacterial DNA release and PCR
  • Enriched samples are prepared and lysed prior to PCR
  • PCR process uses a probe, primers, and template for internal amplification control (IAC)
  • Results displayed directly on the software, then can be reported, stored, and downloaded
  • 96 tests per kit

Kit Contents, Storage, and Preparation

  • Lysis Reagent 1 Tubes (PT0010A) containing lysis reagent 1 and fine white particles
  • Lysis Tube Caps (PT0020A)
  • Proteinase K (PT0050B)
  • SureTect Salmonella PCR Tubes (PT0151A) containing pale yellow pellets
  • PCR Caps (PT0040A)
  • Stored at 2-8°C before use
  • PCR tubes should be brought to room temperature before use

Materials Required (but not provided)

  • Enrichment media (e.g., Buffered Peptone Water (ISO))
  • Other culture formats (e.g., Dry-Bags™)
  • Homogenizer, blender
  • Homogenizer bags
  • Pipettes
  • Microfuge tubes
  • Sample tube rack
  • Culture media
  • Equipment (optional)

Test Procedure

  • Sampling and Enrichment protocols vary depending on the food type.
  • Specific protocols detailed for Raw beef meat, Meat products, Pet food, Dairy products, Powdered infant formula, Seafood and Vegetable products, and Production environment samples.
  • Incubations times and temperatures vary depending on the matrix
  • Samples are processed following the appropriate protocols
  • Positive results confirmed by plating onto Brilliance Salmonella Agar, using a diminishing streak, and incubating at 37°C ±1°C for 24 hours.

Confirmation of Positive Results

  • If not immediately confirmed, store enrichments at 2-8°C for 72 hours.
  • Confirmation by alternate tests, like conventional tests, or Oxoid Salmonella Latex Test/Microbact GNB 24E kit

Quality Control

  • Internal Amplification Control (IAC) targets presence verifies PCR success

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Related Documents

SureTect Salmonella species PCR Assay PDF

Description

This quiz provides an overview of the Thermo Scientific™ SureTect™ Salmonella species PCR Assay PT0100A. It covers the assay's application for detecting Salmonella in food samples, the technology used, and the key features such as result reporting and test kit details. Test your knowledge on this essential tool for food safety and microbiological testing.

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