An integrated microfluidic system using magnetic beads for virus detection.
An integrated system capable of sample pretreatment using antibody-conjugated magnetic beads and one-step reverse transcriptase-polymerase chain reaction (RT-PCR) on a microfluidic system was developed to accelerate the detection of RNA viruses such as dengue virus or enterovirus 71. The targeted virus in the sample was first captured by the specific antibody-conjugated magnetic beads, which were manipulated by micro-electromagnets made of micro-electro-mechanical systems technology. The RNA of the targeted virus then underwent thermal lysis and was reverse-transcripted to cDNA using a microRT-PCR module. The sensitivity to detect dengue virus is around 10-100 PFU, which is equivalent to the commercial RNA extraction kit and a large-scale RT-PCR machine. This microsystem can specifically detect 4 serotypes of dengue virus, as well as enterovirus 71. The specificity was warranted by both antibody and primer. The microfluidic system allows automatic process of sample including mixing, incubation, and reaction. The antibody-conjugated magnetic beads offer sample pretreatment of purification and concentration. The integration of antibody-conjugated magnetic beads into the microfluidic system is promising for fast molecular diagnosis of microorganisms.
Lee
Wan-Chi
WC
Department of Engineering Science, National Cheng Kung University, Tainan 701, Taiwan.
Lien
Kang-Yi
KY
Lee
Gwo-Bin
GB
Lei
Huan-Yao
HY
D003716:Dengue Virus / Q000235:genetics / Q000302:isolation & purification; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification; D008280:Magnetics; D008863:Microspheres; D012327:RNA Virus Infections / Q000175:diagnosis*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*; D014780:Viruses / Q000235:genetics / Q000302:isolation & purification*
D012367:RNA, Viral