Kit per il test dell'influenza aviaria (virus sottotipo H1N1) Kit per il rilevamento con PCR in tempo reale
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Product Name (EN): Avian Influenza Test Kit
Package Size: 50 tests
Price: Contact Our Sales
Stoccaggio: Transport at low temperature; store at -20°C, valid for 12 months
Shipping From: Shanghai, China
For Research Use Only
Avian Influenza Test Kit Overview
This Avian Influenza Test Kit is specifically designed for the qualitative and quantitative detection of Influenza A virus H1N1 subtype RNA using probe-based fluorescent qRT-PCR technology. By targeting conserved genetic regions of the H1N1 subtype, the kit delivers high specificity and sensitivity in detecting avian influenza A virus from biological and environmental samples.
Unlike SYBR Green dye-based PCR, this TaqMan probe-based system uses fluorescence-labeled probes that emit signals only upon hybridization and cleavage during amplification, ensuring superior specificity and lower false-positive rates.
Caratteristiche principali
✅ Highly Specific & Sensitive – Designed based on highly conserved regions of the H1N1 genome; no cross-reaction with non-target viruses
✅ Ready-to-Use System – Requires only RNA template from the user
✅ Optimized Primers & Probes – Ensures efficient amplification and detection
✅ Positive Control Included – Synthetic non-infectious DNA for result validation
✅ Quantitative & Qualitative Applications – Suitable for research, surveillance, and screening use
✅ Compatible with Major qPCR Instruments – Fluorescence detection on FAM channel
Kit Contents (50 tests)
Component Description | Code | Volume & Cap Color |
---|---|---|
qRT-PCR Buffer (probe system) | A | 500 µL |
qRT-PCR Enzyme Mix | B | 100 µL |
PCR Template Dilution Buffer | C | 1 mL |
qRT-PCR Primer Mix for H1N1 | D | 100 µL |
Probe for H1N1 Subtype | E | 50 µL |
Positive Control DNA (1×10⁸ copies/μL) | F | 50 µL |
RNA Release Reagent (trial, extraction-free method) | G | 20 tests |
User Manual | H | 1 copy |
How It Works
The kit detects viral RNA through a one-step reverse transcription PCR reaction. During amplification, the Taq polymerase cleaves the dual-labeled probe, separating the fluorophore from the quencher, which results in a fluorescence signal proportional to the amount of target RNA present.
Fluorescence is measured during each cycle, allowing real-time monitoring of the PCR process.
Principle of Detection
Ct Value (Cycle Threshold): The number of PCR cycles required for fluorescence to exceed the threshold. Lower Ct indicates a higher initial target quantity.
Threshold Setting: Based on 10× standard deviation of fluorescence from the initial 3–15 cycles (background).
Performance & Cut-Off Criteria
Result Type | Criteria |
---|---|
Positivo | Ct ≤ 35 with a typical S-shaped amplification curve |
Suspected Positive | Ct 35–40 → Repeat test recommended |
Negativo | Ct ≥ 40 or no Ct + no amplification curve |
Positive and negative controls must pass quality checks. Ct of negative control ≥40 and positive control Ct ≤30 with exponential amplification.
Reaction Protocol
Reverse Transcription + PCR (20 μL per reaction)
Reagent | Volume |
---|---|
qRT-PCR Buffer | 10 μL |
Enzyme Mix | 2 μL |
Primer Mix | 2 μL |
Probe | 1 μL |
RNA Template or Control | 5 μL |
Total Volume: 20 μL
Thermal Cycling Conditions:
Passo | Temperatura | Tempo |
---|---|---|
Trascrizione inversa | 50°C | 30 min |
Initial Denaturation | 94°C | 10 min |
PCR Cycling × 40 | ||
– Denaturation | 94°C | 15 sec |
– Annealing/Extension | 60°C | 1 min (FAM) |
Quantification (Optional)
Prepare a standard curve using serial 10-fold dilutions (10²–10⁷ copies/μL) of the synthetic DNA control. Plot Ct values against log(concentration) to interpolate RNA copy numbers in unknown samples.
Advantages of Probe-Based PCR Over SYBR Green:
Feature | SYBR Green | Probe-Based (This Kit) |
---|---|---|
Specificity | Binds all dsDNA | Binds only to a specific target sequence |
False Positives Risk | Higher | Lower |
Costo | Lower | Higher (but more reliable) |
Ease of Use | Simpler | Requires probe design |
Intended Use
This product is for research purposes only. It is not approved for clinical diagnostics or therapeutic procedures.
Recensioni
Ancora non ci sono recensioni.