Product Name

HDV (Hepatitis D Virus) Real-Time RT-PCR Detection Kit

Intended Use

For the in vitro qualitative detection of HDV RNA in human serum or plasma. Designed for diagnosis, monitoring, and viral load quantification of Hepatitis D Virus infection.

Technical Parameters:

Principle of the Test

• Real-Time RT-PCR: Targets a conserved region of the HDV genome (e.g., R0 or HDAg gene) using sequence-specific primers and TaqMan probes (e.g., FAM-labeled).

• High Sensitivity & Specificity: No cross-reactivity with HBV, HCV, or other hepatitis viruses.

• Internal Control (IC): Included to monitor RNA extraction and PCR inhibition (e.g., RNase P or synthetic RNA control).

Kit Components

1. RT-PCR Master Mix: Contains reverse transcriptase, hot-start Taq polymerase, dNTPs, and optimized buffer.

2. Primer/Probe Mix: HDV-specific oligonucleotides with fluorescent probe (e.g., FAM for HDV, HEX/VIC for IC).

3. Controls:

   • Positive Control: Synthetic HDV RNA or inactivated virus.

   • Negative Control: Nuclease-free water.

   • Internal Control (if included).

4. Other Reagents: DNase/RNase-free water, reaction tubes (if provided).

Workflow

1. Sample Preparation: Viral RNA extraction (compatible with QIAamp, MagMAX, or automated systems).

2. PCR Setup: Mix extracted RNA with master mix and primers/probes.

3. Amplification:

   • Reverse Transcription: 50°C for 10–15 min.

   • PCR Activation: 95°C for 2–5 min.

   • Cycling: 40–45 cycles of 95°C (10 sec) → 60°C (30–60 sec, fluorescence read).

4. Analysis:

   • Quantitative: Standard curve for viral load (IU/mL or copies/mL).

   • Qualitative: Ct value cut-off for positive/negative results.

Performance Characteristics

• Sensitivity: ≤10–50 IU/mL (95% LOD).

• Linear Range: 10²–10⁸ IU/mL (quantitative kits).

• Specificity: No cross-reactivity with HBV, HCV, HIV, or other common pathogens.

• Precision: CV <5% for intra- and inter-assay variability.

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