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Volume 15
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10.3390/pathogens15060600
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Article

Evaluation of IndiMix JOE with Intype IC-RNA as an Alternative to AgPath-ID for Influenza A Virus Detection in Avian and Bovine Samples

by
Andie Hach
1,
Anne Vandenburg-Carroll
1,
Douglas G. Marthaler
2,
Stephen Vadia
3,
Qirui Zhang
3,
Melanie Prarat
3 and
Ailam Lim
1,4,*
1
Wisconsin Veterinary Diagnostic Laboratory, University of Wisconsin–Madison, Madison, WI 53706, USA
2
INDICAL BIOSCIENCE, 04103 Leipzig, Germany
3
Ohio Department of Agriculture, Animal Disease Diagnostic Laboratory, Reynoldsburg, OH 43068, USA
4
Department of Pathobiological Sciences, University of Wisconsin–Madison, Madison, WI 53706, USA
*
Author to whom correspondence should be addressed.
Pathogens 2026, 15(6), 600; https://doi.org/10.3390/pathogens15060600
Submission received: 18 May 2026 / Revised: 23 May 2026 / Accepted: 26 May 2026 / Published: 3 June 2026
(This article belongs to the Section Viral Pathogens)
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Abstract

This method comparison demonstrated equivalent analytical and diagnostic performance of IndiMix JOE with intype IC-RNA as an alternative real-time polymerase chain reaction (RT-PCR) chemistry compared to the National Animal Health Laboratory Network (NAHLN) influenza A virus (IAV) RT-PCR surveillance assay using AgPath-ID One-Step RT-PCR Reagents in avian swabs and tissues and bovine milk. In avian samples, IndiMix JOE with intype IC-RNA using a fast reduced-volume protocol had comparable results to the NAHLN reference RT-PCR with and without intype IC-RNA using standard NAHLN thermocycling conditions. A six-way comparison of RT-PCR chemistries and exogenous internal controls in milk samples illustrated equivalent mean CT values (ANOVA, p = 0.9938). Additional experiments in avian swabs and tissues, bovine milk and semen samples resulted in comparable analytical sensitivity in limits of detection (LOD), linearity (R2 > 0.977), and PCR efficiencies, and no significant differences in mean CT values (ANOVA, p > 0.05). Diagnostic performance showed 100% agreement across field and spiked sample matrices compared to the NAHLN reference method. The study supports the use of IndiMix JOE and intype IC-RNA as an alternative, with practical improvements relevant to surveillance workflows including enhanced testing flexibility, efficiency, outbreak response capacity, and reduced time to results.
Keywords: influenza A virus surveillance; highly pathogenic avian influenza; exogeneous internal controls; diagnostic method comparison; limit of detection; analytical sensitivity; RT-PCR; PCR inhibition; outbreak response; IndiMix JOE; intype IC-RNA; AgPath-ID One-Step RT-PCR Reagents; bovine milk; bovine semen influenza A virus surveillance; highly pathogenic avian influenza; exogeneous internal controls; diagnostic method comparison; limit of detection; analytical sensitivity; RT-PCR; PCR inhibition; outbreak response; IndiMix JOE; intype IC-RNA; AgPath-ID One-Step RT-PCR Reagents; bovine milk; bovine semen

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MDPI and ACS Style

Hach, A.; Vandenburg-Carroll, A.; Marthaler, D.G.; Vadia, S.; Zhang, Q.; Prarat, M.; Lim, A. Evaluation of IndiMix JOE with Intype IC-RNA as an Alternative to AgPath-ID for Influenza A Virus Detection in Avian and Bovine Samples. Pathogens 2026, 15, 600. https://doi.org/10.3390/pathogens15060600

AMA Style

Hach A, Vandenburg-Carroll A, Marthaler DG, Vadia S, Zhang Q, Prarat M, Lim A. Evaluation of IndiMix JOE with Intype IC-RNA as an Alternative to AgPath-ID for Influenza A Virus Detection in Avian and Bovine Samples. Pathogens. 2026; 15(6):600. https://doi.org/10.3390/pathogens15060600

Chicago/Turabian Style

Hach, Andie, Anne Vandenburg-Carroll, Douglas G. Marthaler, Stephen Vadia, Qirui Zhang, Melanie Prarat, and Ailam Lim. 2026. "Evaluation of IndiMix JOE with Intype IC-RNA as an Alternative to AgPath-ID for Influenza A Virus Detection in Avian and Bovine Samples" Pathogens 15, no. 6: 600. https://doi.org/10.3390/pathogens15060600

APA Style

Hach, A., Vandenburg-Carroll, A., Marthaler, D. G., Vadia, S., Zhang, Q., Prarat, M., & Lim, A. (2026). Evaluation of IndiMix JOE with Intype IC-RNA as an Alternative to AgPath-ID for Influenza A Virus Detection in Avian and Bovine Samples. Pathogens, 15(6), 600. https://doi.org/10.3390/pathogens15060600

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