Kit de detecção de ácido nucleico de Salmonella UNG (PCR em tempo real 50T/100T)

Catalog Number: AP2409004
Specifications: 50 Tests / 100 Tests
Armazenamento: -20°C

Product Description

The Salmonella Nucleic Acid Detection Kit is a real-time PCR assay designed for the rapid, specific, and sensitive detection of Salmonella DNA in a variety of sample types, including stool, water, tissue, and food. Salmonella is a major foodborne pathogen commonly transmitted through contaminated eggs, poultry, and meat.

This kit uses fluorescent probe-based quantitative PCR (qPCR) technology to amplify a specific fragment of the Salmonella genome. Results are determined by Ct (cycle threshold) values or via standard curve quantification to estimate the bacterial load in the sample.

Key Features

• ✅ High specificity and sensitivity using the TaqMan fluorescence probe method

• ✅ Compatible with multiple real-time PCR systems (FAM channel)

• ✅ Includes positive and negative controls for result validation

• ✅ UNG enzyme included to eliminate carryover contamination

• ✅ Suitable for food safety monitoring, animal disease diagnostics, and research

Kit Components

Sample Types & Preparation

• Swabs (Oral/Cloacal): For birds and poultry, insert a sterile swab into the throat and cloaca, rotate gently, then transfer to a sterile centrifuge tube.

• Whole Blood: Use EDTA anticoagulant only. Do not use heparin. Fresh blood is recommended. Store at 2–8°C for up to 7 days, or at -20°C for up to 3 months.

• Tissue Samples: Use ≤100 mg of liver or other tissues, homogenize with 200–500 μL sterile water.

Note: Pre-enrichment is required based on GB 4789.4-2016 standards or equivalent. Suggested enrichment in BPW broth (1:10 w/v), followed by incubation at 36 ± 1°C for 8–18 hours.

Please use DN/RNA Extraction Kit (Cat. No. DP201) or an equivalent method to extract nucleic acids. Extracted DNA should be kept on ice and tested promptly (≤7 days at 4°C or ≤6 months at -20°C).

PCR Protocol

1. Reaction Mix Preparation:

   • Thaw all reagents at room temperature and centrifuge briefly.

   • Dispense 23 μL of Salmonella PCR mix into each PCR tube.

   • Add 2 μL of sample DNA, positive control, or negative control to each tube, respectively.

   • Final volume: 25 μL per reaction.

2. PCR Program:

   • UNG treatment: 50°C for 2 min

   • Initial denaturation: 95°C for 5 min

   • 40 cycles:

     • Denaturation: 95°C for 10 sec

     • Annealing/Extension: 60°C for 35 sec (FAM signal collected)

3. Fluorescence Channel:

   • FAM (For ABI series and other qPCR instruments)

   • Add ROX reference dye if required by the instrument.

Result Interpretation

Note: Suspect results may be caused by inhibitors (e.g., alcohol, anticoagulants) or low DNA yield. If aerosol contamination is ruled out and Ct <35 upon retesting, the result is considered positive.

Quality Control Criteria

• Positive Control: FAM Ct <28 with S-shaped amplification curve

• Negative Control: No Ct or Ct ≥40 without S-curve

• Otherwise, results are considered invalid and retesting is required.

Precautions

1. Zoned Workflow: Use physical separation for pre-PCR, reagent setup, and amplification areas. Wear protective gear and change gloves between zones.

2. Avoid Contamination: Delay tube opening post-PCR to avoid aerosol generation. UV or chemical sterilization of equipment is strongly recommended.

3. Freeze/Thaw Cycles: Avoid repeated freeze-thaw of reagents. Fully thaw and gently mix before use.

4. Research Use Only: This kit is for scientific research and food/environmental safety screening only. Not for clinical diagnostic use.

Storage and Shelf Life

• Store at -20°C

• Valid for at least 12 months if unopened and properly stored

• Protect from light and repeated freeze-thaw cycles