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Cat. No.: FPVFL-25 (for 25T)

Cat. No.: FPVFL-100 (for 100T)

Description

Feline Panleukopenia Virus (FPV) Fluorescence LAMP Assay Kit is designed to detect feline parvovirus DNA using Loop Mediated Isothermal Amplification (LAMP) and to determine infection through fluorescence curves on a quantitative PCR instrument. This kit incorporates dU and UDG enzyme to prevent false positives from contamination of amplification products, enabling rapid, efficient, and highly sensitive detection of feline parvovirus.

Feline Panleukopenia Virus (FPV), also known as feline distemper virus or feline infectious enteritis virus, is a single-stranded DNA virus in the Parvoviridae family. Under an electron microscope, FPV appears round or hexagonal without an envelope. The genome is approximately 5200 nucleotides long and contains two open reading frames (ORFs) that encode structural proteins (VP1 and VP2) and non-structural proteins (NS1 and NS2). Infected animals experience a sharp decline in white blood cells, severe fever, vomiting, and diarrhea. The virus primarily affects young cats, is highly contagious among all felines with a high mortality rate, and spreads rapidly through the feces, urine, and vomit of infected animals. It can be vertically transmitted during pregnancy, leading to cerebellar hypoplasia in fetuses. Early and rapid diagnosis of FPV is crucial to isolate infected cats and provide targeted treatment to reduce morbidity and mortality.

The main symptoms of FPV infection include fever, vomiting, and diarrhea, which can easily be confused with other gastrointestinal diseases, making diagnosis based solely on clinical symptoms difficult. Current clinical diagnostic methods for FPV infection include electron microscopy (EM), virus isolation (VI), latex agglutination test (LAT), hemagglutination (HA), ELISA, and PCR analysis. While electron microscopy and virus isolation are specific and sensitive, they are expensive and time-consuming in clinical use. Latex agglutination is fast but lacks specificity. Hemagglutination or serum hemagglutination inhibition tests can produce false negatives due to weak hemagglutination activity of FPV and stringent operating conditions. Although ELISA and PCR are widely used for their high sensitivity and specificity, they require specialized equipment and technical expertise, limiting their applicability.

Isothermal amplification technology is an in vitro nucleic acid amplification technique that maintains a constant temperature throughout the reaction, enabling rapid nucleic acid amplification by adding enzymes with different activities and their specific primers. The kit uses Loop-Mediated Isothermal Amplification (LAMP) technology, which designs 4-6 specific primers targeting six regions of the target gene. Bst DNA Polymerase initiates DNA synthesis, forming dumbbell-shaped complementary chains that further undergo continuous strand displacement to enter the cyclic amplification stage. The final amplified product is a mixture of DNA with various stem-loop structures and cauliflower-like structures. LAMP requires only 30-60 minutes of incubation at isothermal conditions (e.g., 60-65ºC) to complete nucleic acid amplification. Compared to conventional PCR, LAMP does not require template denaturation, temperature cycling, electrophoresis, or UV observation, offering simplicity, rapidity, high sensitivity, and strong specificity. The kit targets the most conserved region of the FPV VP2 gene, designing specific primers compatible with most FPV variants.

Components

• LAMP Fluorescence Master Mix with UDG (2X)
• LAMP Primer Mix (10X)
• Bst DNA Polymerase
• Positive Control
• Nuclease-free Water

Features

• This kit offers high sensitivity and a short reaction time. Its sensitivity is 2-5 times higher than traditional PCR methods, with a detection limit of approximately 20-500 copies/μl using positive plasmid calculation, and it only requires 45 minutes to complete the detection reaction.
• The kit provides strong specificity. It uses six specific primers designed based on the conserved sequences of feline panleukopenia virus, ensuring no cross-reactivity with the DNA of other organisms.
• The kit uses anti-contamination technology to effectively avoid contamination. It incorporates dU and heat-labile UDG enzyme, allowing dU to be incorporated into the amplification products and effectively eliminating contamination issues from the amplification process by incubating at 37ºC for 5-15 minutes before isothermal amplification. During isothermal amplification, the heat-labile UDG enzyme is inactivated, thus not interfering with the subsequent detection.
• The kit includes a Positive Control to ensure the kit is functioning correctly. It is recommended to include a positive control in each test.