title: Integrated microfluidic reverse transcription-polymerase chain reaction for rapid detection of food- or waterborne pathogenic rotavirus.
abstract: The development of microfluidic tools for nucleic acid analysis has become a burgeoning area of research during the postgenome era. Here we have developed a microfluidic device that integrates reverse transcription (RT) and polymerase chain reaction (PCR) with online fluorescence detection to realize a rapid detection system for performing both genetic amplification and product analysis. The microfluidic device mainly comprises a grooved copper heating block for RT and a heated cylinder for amplification. To expedite the analysis process, we combined the continuous-flow PCR with an online fluorescence detection system that allows analysis of amplification products within 1 min. Rotaviruses are worldwide enteric pathogens in humans and animals responsible for a significant burden of disease through person-to-person transmission and exposure to contaminated foods and water. In this study, rotavirus from stool specimens was successfully amplified and detected using the RT-PCR microfluidic system within 1 h, and the limit of detection of the RNA concentration was estimated to be 3.6×10(4) copies μl(-1). Compared with a large-scale apparatus, the integrated microfluidic system presented here can perform rapid nucleic acid amplification and analysis, possibly making it a crucial platform for future diagnosis application.
authors.affiliation: MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, People's Republic of China.
authors.forename: Yuyuan
authors.initials: Y
authors.lastname: Li
authors.forename: Chunsun
authors.initials: C
authors.lastname: Zhang
authors.forename: Da
authors.initials: D
authors.lastname: Xing
mesh_terms: D005243:Feces / Q000821:virology; D005456:Fluorescent Dyes / Q000737:chemistry; D005502:Food / Q000821:virology; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000175:diagnosis / Q000821:virology; D014871:Water Microbiology
chemical_list: D005456:Fluorescent Dyes; D012367:RNA, Viral