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 DescriptionCodeVolume & Cap Color
qRT-PCR Buffer (probe system)A500 µL
qRT-PCR Enzyme MixB100 µL
PCR Template Dilution BufferC1 mL
qRT-PCR Primer Mix for H1N1D100 µL
Probe for H1N1 SubtypeE50 µL
Positive Control DNA (1×10⁸ copies/μL)F50 µL
RNA Release Reagent (trial, extraction-free method)G20 tests
User ManualH1 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 TypeCriteria
PositivoCt ≤ 35 with a typical S-shaped amplification curve
Suspected PositiveCt 35–40 → Repeat test recommended
NegativoCt ≥ 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)

ReagentVolume
qRT-PCR Buffer10 μL
Enzyme Mix2 μL
Primer Mix2 μL
Probe1 μL
RNA Template or Control5 μL
  • Total Volume: 20 μL

  • Thermal Cycling Conditions:

PassoTemperaturaTempo
Trascrizione inversa50°C30 min
Initial Denaturation94°C10 min
PCR Cycling × 40
– Denaturation94°C15 sec
– Annealing/Extension60°C1 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:

FeatureSYBR GreenProbe-Based (This Kit)
SpecificityBinds all dsDNABinds only to a specific target sequence
False Positives RiskHigherLower
CostoLowerHigher (but more reliable)
Ease of UseSimplerRequires probe design

Intended Use

This product is for research purposes only. It is not approved for clinical diagnostics or therapeutic procedures.

Recensioni

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