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?

Fluorescent dyes for staining Signup and view all the answers

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

37°C ±1°C, 41.5°C ±1°C Signup and view all the answers

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

It enhances the probe-template DNA bond. Signup and view all the answers

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

1 hour and 20 minutes Signup and view all the answers

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

Oxoid Brilliance™ Salmonella Agar Signup and view all the answers

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

Conduct a full evaluation of the enrichment procedure Signup and view all the answers

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

Compliance with Good Laboratory Practices Signup and view all the answers

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

Oxoid Cocoa Samples Recovery (CSR) Broth Signup and view all the answers

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

20mg/l of novobiocin Signup and view all the answers

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

Lysis Tube Rack Signup and view all the answers

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

SureTect PikoReal Real-Time PCR Instrument Signup and view all the answers

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

Windows 7 Signup and view all the answers

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

Add 1.5 litres of pre-warmed mTSB at 41.5°C. Signup and view all the answers

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

It should be diluted 1:5 using sterile CSR Broth or Peptone Salt solution. Signup and view all the answers

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

37°C ±1°C. Signup and view all the answers

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

Pre-moisten with a neutralizing broth like Dey-Engley Broth. Signup and view all the answers

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

Up to 2 hours. Signup and view all the answers

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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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.