Four Bovine Serum Virus Probe qPCR Kit

Description

This kit is designed for the qualitative detection of nucleic acids from four major bovine serum viruses: Bovine Leukemia Virus (BLV), Foot-and-Mouth Disease Virus (FMDV), Rabies Virus (RBV), and Bluetongue Virus (BTV). It provides rapid, sensitive, and reliable results for veterinary research and diagnostic applications.

Principle

The assay is based on multiplex real-time PCR technology with target-specific primers and fluorescent probes directed against conserved genomic regions of the viruses. The 5′ nuclease (TaqMan) assay generates fluorescence upon probe cleavage during amplification. An internal control is included to monitor nucleic acid extraction and amplification efficiency, preventing false negatives. The dU–UNG system minimizes carryover contamination and reduces false positives.

Kit Components

• Nucleic acid extraction and purification kit (magnetic bead-based reagents)

• Ready-to-use PCR mix in strip tubes (primers, probes, Mg²⁺, dNTPs, reverse transcriptase, RNase inhibitor, UNG, Taq polymerase)

• Negative control (800 µL bovine genomic DNA)

• Positive control (1.0 mL plasmid/pseudovirus with target sequences, non-infectious)

Storage

• Store at –20 ± 5 °C, protected from light

• Shelf life: 12 months

• Avoid >5 freeze–thaw cycles

Compatible Instruments

• ABI 7500

• SLAN-96P

• AutoPlex-Mini

Sample Requirements

• Sample type: Bovine serum

• Storage: ≤12 hours at 2–8 °C; long-term at –70 °C

• Transport: Ice pack (≤1 day); dry ice (long distance)

Testing Procedure

AutoPlex-Mini (Automated workflow): Load 200–300 µL serum for fully automated extraction and amplification.

Manual extraction (ABI 7500/SLAN-96P): Use 200 µL serum, add 10 µL nucleic acid extract to PCR reaction, and follow standard thermal cycling parameters.

Performance Characteristics

• Sensitivity: 500 copies/mL for each target virus

• Specificity: No cross-reactivity with other bovine viruses

• Precision: CV ≤ 5.0% (n=10)

Quality Control

• Negative control: Internal control Ct ≤35; no amplification in target channels

• Positive control: S-shaped curve with Ct 25–38

Interpretation of Results

• Positive: Ct ≤38 with amplification curve

• Negative: No Ct or Ct >38 (valid only if internal control amplifies correctly)