title: Microchip capillary electrophoresis with laser-induced fluorescence combined with one-step duplex reverse-transcription polymerase chain reaction for the rapid detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens.
abstract: Enterovirus 71 and Coxsackievirus A16 are the main pathogens causing hand-foot-mouth disease. In this paper, microchip capillary electrophoresis with laser-induced fluorescence combined with one-step duplex reverse transcript-polymerase chain reaction has been developed for the detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens. The specific reverse transcription-polymerase chain reaction amplicons labeled with SYBR Orange were separated by microchip capillary electrophoresis and detected by laser induced fluorescence detector within 7 min. The intraday and interday relative standard deviation of migration time for DNA Marker was in the range of 1.36-2.94 and 2.78-3.96%, respectively. The detection limits were as low as 2.06 × 10(3) copies/mL for Enterovirus 71 and 5 × 10(3) copies/mL for Coxsackievirus A16. No cross-reactivity was observed with rotavirus, astrovirus, norovirus, and adenovirus, which showed good specificity of the method. This assay was validated using 100 throat swab specimens that were detected by real-time reverse-transcript polymerase chain reaction in parallel and the two methods produced the same results. This study provided a rapid, sensitive and specific method for the detection of Enterovirus 71 and Coxsackievirus A16, which make a contribution to significant time and cost saving for the identification and treatment of patients.
authors.affiliation: West China School of Public Health, Sichuan University, Chengdu, China.
authors.forename: Ruan
authors.initials: R
authors.lastname: Jia
authors.affiliation: West China School of Public Health, Sichuan University, Chengdu, China.|Sichuan Province Key Laboratory of Food Safety Monitoring and Risk Assessment, Chengdu, China.
authors.forename: Sun
authors.initials: S
authors.lastname: Chengjun
authors.affiliation: Department of Microbiological Experiment, Chengdu Center for Disease Control and Prevention, Chengdu, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Heng
authors.affiliation: West China School of Public Health, Sichuan University, Chengdu, China.
authors.forename: Zhou
authors.initials: Z
authors.lastname: Chen
authors.affiliation: West China School of Public Health, Sichuan University, Chengdu, China.
authors.forename: Li
authors.initials: L
authors.lastname: Yuanqian
authors.affiliation: West China School of Public Health, Sichuan University, Chengdu, China.|Sichuan Province Key Laboratory of Food Safety Monitoring and Risk Assessment, Chengdu, China.
authors.forename: Li
authors.initials: L
authors.lastname: Yongxin
mesh_terms: D004279:DNA, Viral / Q000032:analysis; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D004770:Enterovirus / Q000302:isolation & purification*; D006232:Hand, Foot and Mouth Disease / Q000175:diagnosis / Q000821:virology; D006801:Humans; D057230:Limit of Detection; D010614:Pharynx / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods*
chemical_list: D004279:DNA, Viral
keywords: Coxsackievirus A16; Enterovirus 71; Laser-induced fluorescence; Microchip capillary electrophoresis; Throat swab specimens