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Fecal Bacteriology PCR Kit
BIOSYNEX AMPLIQUICK Fecal Bacteriology detects 15 bacterial agents of gastroenteritis in a DNA extract from stool, by real-time PCR in five master mixes with detection in the FAM, HEX, Texas Red and Cy5 channels. Every sample carries two controls: the CIEZ internal control, which shows whether the PCR was inhibited, and a procedural control based on a Drosophila Rp49 sequence, which shows whether extraction and purification worked. The kit was evaluated by the French national reference centre for Campylobacter and Helicobacter.
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This kit in detail
- 15 bacterial targets across five ready-to-use master mixes
- Internal amplification control and procedural control for every patient
- Limit of detection 95 per cent between 0.2 and 2.2 copies per µl for the strains tested
- Stool via AMPLIQUICK Fecal Pretreatment or a Copan FecalSwab
- Amplification in about 1 hour 15 minutes
- Plate format with 60 reactions, or bulk tubes with 96 reactions per master mix
BIOSYNEX AMPLIQUICK Fecal Bacteriology is a real-time PCR test for the qualitative detection of 15 bacterial agents commonly associated with gastroenteritis, as an aid to diagnosis on a purified DNA extract from stool.
Targets and layout
The targets are divided over five master mixes, read in the FAM, HEX and Texas Red channels with the control in Cy5. Master mix A covers Campylobacter species (16S, lpxA and gyrB), Yersinia enterocolitica and Y. pseudotuberculosis (yst and ail) and Salmonella species (invA). Master mix B covers hypervirulent Clostridioides difficile (tcdC delta117), C. difficile toxin B and binary toxin (tcdB and CDT) and Aeromonas species (gyrB). The remaining master mixes complete the panel of 15 targets, with the procedural control read in master mix E.
Two limitations worth knowing
Yersinia enterocolitica and Y. pseudotuberculosis are read in the same channel and are therefore not distinguished from one another, and the same applies to C. difficile toxin B and the binary toxin strains. Liquid stool can give a false negative through dilution; the manufacturer then advises direct extraction from 200 µl.
Performance
The 95 per cent limit of detection lies between 0.2 and 2.2 copies per µl for the strains tested, which is 1.2 to 11 copies per reaction. Reported values include 0.245 copies per µl for Campylobacter coli, 0.579 for C. jejuni, 1.264 for C. upsaliensis, 1.978 for C. fetus, 1.167 for Y. pseudotuberculosis and 2.221 for Y. enterocolitica.
Sample and instruments
Stool collected with the AMPLIQUICK Fecal Pretreatment kit (REF 3150065) or with a Copan FecalSwab. Five µl of extraction eluate goes into a reaction volume of 20 µl. Validated extraction systems are the Macherey-Nagel NucleoMag Dx Pathogen with MagnetaPure 32 and the QIAGEN QIAamp Fast DNA Stool Mini kit; validated thermocyclers are the Bio-Rad CFX96 Touch and CFX96 OPUS, the Bioer QuantGene 9600 and the Applied Biosystems QuantStudio 5. The kit is stored at or below -20 °C. Three formats are available: two plate formats with 60 reactions (REF 3150046_SEC01 with low-profile wells and SEC02 with high-profile wells) and a bulk format with 96 reactions per master mix (REF 3150046_BLK96).
- Master Mixes (A, B, C, D, E): Ready-to-use solutions targeting specific bacterial genes and toxins.
- Positive Control: Validates the PCR amplification step.
- Negative Control: Ensures no contamination in the process.
- Control in Process: Monitors sample quality and extraction efficiency.
- Master Mix Control: Verifies the performance of master mixes.
- Optical Cap Strips: Seal PCR plates to prevent contamination.
- Prefilled Microplates (Optional): Preloaded for easy and high-throughput testing.
Datasheets and regulatory documentation
Everything your quality team needs for evaluation and procurement.
Instructions for Use
Frequently paired with this kit
Other assays and instruments lab teams add alongside the Fecal Bacteriology PCR Kit.
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