Suitability of Commercial FeLV Nucleic Acid Detection Kits for Small Lab Environments
Feline leukemia retrovirus (FeLV) causes a chronic infectious disease manifested by profound anemia, malignancies, and immunosuppression. If present in multi-cat environments, FeLV has an impact on survival and quality-of-life, therefore, accurate and early detection is imperative to prevent the virus from spreading to other cats.
Comparison of two commercially available molecular kits for the detection of FeLV RNA and DNA using several well-known nucleic acid extraction kits.
Molecular kits, Biogal PCRun® and PrimerDesign were tested and compared to each other to define the sensitivity, specificity, and suitability for use in small laboratory settings. DNA and/or RNA (n=121) were purified from three different retrospective sample archives (buffy coat and plasma) collected in Switzerland (Group 1), Portugal (Group 2), and USA (Group 3) employing extraction kits from 2 different companies (Qiagen and Zymo). Final results were compared to the Cq values or SNAP® FeLV/FIV generated by the laboratory source.
PCRun® results correlated better with TaqMan® (sensitivity 100%/specificity 84%) than with SNAP®.
Extraction methods affected final results. Zymo preps generated DNA with lower inhibition of the PCRun® reactions (Sensitivity: Zymo 92–97%; Qiagen 74%). Zymo produced poorer quality RNA extracts compared to Qiagen (Sensitivity 89.5–91.5%).
Results obtained with PCRun® were 100% complementary to the TaqMan® and matched closely to the viremic stages of the subjects.
PCRun® is a point-of-care test, while PrimerDesign® is a Taqman® RealTime PCR. The hands-on application and accuracy of PCRun® makes it highly suitable for screening of FeLV in laboratories with minimal equipment.