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 and Stability

Four Bovine Serum Virus Probe qPCR Kit (PLEXD02A) store at -20°C;
Nucleic acid extraction or purification reagents (PLEXD02B)store at room temperature; do not freeze
Shelf life: 12 months
Avoid >5 freeze–thaw cycles

Compatible Instruments

ABI 7500
SLAN-96P
AutoPlex-Mini

Sample Requirements

Sample type: Bovine serum
Storage: Short-term (≤12 hrs): 2–8°C. Long-term: -70°C (avoid freeze-thaw).
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)

Compatible Devices

AutoPlex-12

AutoPlex-12 Mini

Documents

User Manual

Four Bovine Serum Virus Probe qPCR Kit