Development of multiplex nucleic acid sequence-based amplification for detection of human respiratory tract viruses

A group of common lower respiratory tract infections, influenza A, influenza B, human parainfluenza virus 1-4 (HPIV1-4), respiratory syncytial virus (RSV), rubella virus (RV) and Coxsackie virus (CSV), were selected for the development of a multiplex nucleic acid sequence-based amplification (NASBA) assay. Quantifiable measurement utilizing an enzyme-linked oligonucleotide capture (EOC) optical detection method, which was described previously, alleviated the requirement of specialized instrumentation that is commonly used in other molecular techniques. Multiplex NASBA-EOC provided rapid and specific detection of a single virus from a multiplexed group, reducing laboratory testing time and enabling high throughput screening. The uniquely designed primers and probes proved to be highly sensitive and specific, exemplifying the robustness of the multiplex NASBA-EOC technique.