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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
Author(s) -
Jia Ruan,
Chengjun Sun,
Heng Chen,
Chen Zhou,
Yuanqian Li,
Yongxin Li
Publication year - 2015
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201500165
Subject(s) - capillary electrophoresis , enterovirus , amplicon , coxsackievirus , polymerase chain reaction , virology , reverse transcription polymerase chain reaction , microbiology and biotechnology , real time polymerase chain reaction , chemistry , biology , virus , biochemistry , gene , messenger rna
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.