Abstract
Background
Rapid diagnosis is critical for the early detection and control of transboundary animal diseases such as African Swine Fever (ASF) and Avian Influenza (AI), particularly because it enables timely containment measures at the farm level. In low-resource settings where sample transport, cold chain maintenance, and access to centralized laboratories are challenging, portable qPCR systems offer a clear operational advantage by enabling on-site or near-site testing thus reducing diagnostic turnaround and response time. This study evaluated three qPCR platforms for detecting African Swine Fever Virus (ASFV) and Avian Influenza Virus (AIV): two laboratory-based systems, CFX96 Touch (Bio-Rad) and MIC (BioMolecular Systems), and the portable qPCR platform Franklin by Biomemex. Seventy-two ASF virus samples representing genotypes I, II, IX, XIV, XXIII, and a genotype I/II recombinant from eleven countries were tested on two laboratory platforms (CFX96, MIC) and one rapid platform (Franklin) alongside 50 AI virus samples (H5N1, H9N2) from seven countries.Results
Both laboratory platforms achieved consistent results. Franklin required protocol optimization when the WOAH-recommended protocol produced predominantly false-negative results, but adapted amplification conditions restored performance. Limits of detection per reaction calculated by probit analysis were: CFX96 21.28 (14.85-68.67), MIC 32.76 (21.12-96.11), Franklin with lyophilized reagents 4.12 (2.71-14.97), and Franklin with adapted protocol 6.85 (4.33-20.11). After adaptation, all platforms exhibited high diagnostic sensitivity and specificity for ASFV detection. For the AIV detection protocol, false negative results were observed on MIC and Franklin platforms compared to the CFX96.Conclusions
The laboratory platforms tested in this study (CFX96 and MIC) provided robust high-throughput capabilities, while the portable system (Franklin) offered superior sensitivity but required protocol adaptation of established WOAH-recommended procedures.