Microfluidic device for rapid detection of cytomegalovirus (CMV) by sequence-specific hybridization of PCR-amplified CMV-DNA

This paper reports rapid detection of human CMV by using a microfluidic device fabricated on plastic chip. The method employs post PCR product analysis by sequence-specific hybridization between amplified CMV-DNA target and complementary PNA probe in microchannel for specific detection of CMV. The PCR product solution and PNA probe solution flowed simultaneously along the microchannel forming a laminar flow at the straight channel. Hybridization of PCR amplified target DNA and fluorescently labeled peptide nucleic acid (PNA) probe occurred at the interface of the laminar flow. Secondary laminar flow, on the other hand, is formed at the curving part of the microchannel allowing separation of DNA hybrids. Hybridized DNA were detected by laser induced fluorescence microscopy. Collectively, these features allowed identification of PCR amplified CMV-DNA. Compared to the conventional pp65 antigenemia test, microfluidic device is found to be more sensitive in detecting low-level viremia