Enhancing Diagnostic Resilience: Evaluation of Extraction Platforms and IndiMag Pathogen Kits for Rapid Animal Disease Detection

The United States is facing outbreaks of highly pathogenic avian influenza H5N1 in birds and dairy cattle, along with threats of African swine fever, classical swine fever, and foot-and-mouth disease. While the National Animal Health Laboratory Network (NAHLN) depends on high-throughput testing, the KingFisher Duo Prime, IndiMag 48s, and IndiMag 2 are viable alternatives to aid in outbreak assessments. This study evaluates extraction platforms and the IndiMag Pathogen Kit for detecting the previous listed pathogens. Samples and reference materials were extracted using the MagMAX Viral RNA Isolation Kit, MagMAX CORE Nucleic Acid Purification Kit, and IndiMag Pathogen Kit. Real-time RT-PCR was performed following NAHLN protocols to assess analytical and diagnostic performance. Comparable limits of detection across extraction chemistries, instrumentation, and pathogens were demonstrated, with PCR efficiency ranging between 82.5% and 107.6%. The precision variability was low, with the coefficient of variation ranging from 0.16% to 1.76%. Diagnostic sensitivity and specificity were 100%, with a kappa coefficient of 1.0, indicating strong agreement between methods. These findings support the KingFisher Duo Prime, IndiMag 48s, IndiMag 2, and IndiMag Pathogen Kits as reliable options for NAHLN-approved testing, increasing equipment and reagent alternatives to enhance diagnostic resilience and improve response capabilities to emerging animal health threats.