Chromatographia

, Volume 78, Issue 17–18, pp 1191–1199 | Cite as

Capillary Electrophoresis-Based Detection for Foodborne Enteroviruses in Vegetable Samples

  • Jia Ruan
  • Cheng Jun Sun
  • Fan Chen
  • Chen Zhou
  • Yuan Qian Li
  • Ya Pan Liu
  • Yong Xin Li
Original
First Online:
Received:
Revised:
Accepted:

Abstract

Foodborne enteroviruses such as norovirus, rotavirus and astrovirus can cause gastroenteritis in children under the age of five and infants. In this paper, a novel and ultrasensitive method, combining reverse transcription polymerase chain reaction (RT-PCR) with capillary electrophoresis laser-induced fluorescence (CE-LIF), is proposed to detect three viruses in vegetable samples. The viruses, including norovirus, rotavirus and astrovirus in vegetables, were eluted using glycine buffer (pH 9.5), and concentrated by 15 % polyethylene glycol 6000 (PEG 6000). The primers, targeting the specific and conservative sequences of nucleic acids of the viruses, were synthesized and used in RT-PCR reaction. The amplification products were labeled with highly sensitive SYBR Gold, then separated by capillary electrophoresis and detected by a laser-induced fluorescence detector within 16 min. The intraday and interday relative standard deviation of migration time for the DNA Marker was in the range of 1.09–1.30 and 1.77–2.60 %, respectively, while the detection limits of the three viruses were as low as 1.33 × 102 copies mL−1 for norovirus, 1.86 × 102 copies mL−1 for rotavirus, and 1.40 × 102 copies mL−1 for astrovirus. Meanwhile, the results of homology analysis and specificity experiments showed that the method had good specificity. A fried pickled mustard tuber tested positive for norovirus by the proposed method in an outbreak which happened recently in Chengdu. This protocol demonstrated the possibility for rapid, sensitive and specific detection of foodborne enteric viruses in vegetable samples.

Keywords

Capillary electrophoresis-laser induced fluorescence (CE-LIF) Reverse transcription polymerase chain reaction (RT-PCR) SYBR Gold Enteric virus Vegetable samples 
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Notes

Acknowledgment

This work was supported by grants from Youth Foundation of National Natural Science of China (No. 81102162).

Conflict of interest

We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work and that there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in the manuscript entitled “Capillary Electrophoresis-based Detection for Foodborne Enteroviruses in Vegetable Samples”.

Supplementary material

10337_2015_2931_MOESM1_ESM.tif (214 kb)
Supplementary material 1 (TIFF 214 kb). Figure 1 SuppInfo. The sensitivity of norovirus, rotavirus and astrovirus and the electrophoretogram of Gene RulerTM Low Range DNA Ladder.The figures indicate the sizes of DNA fragments:1.25 bp 2.50 bp 3.75 bp 4.100 bp 5.150 bp 6.200 bp 7.300 bp 8.400 bp 9.500 bp 10.700 bp
10337_2015_2931_MOESM2_ESM.tif (366 kb)
Supplementary material 2 (TIFF 365 kb). Figure 2 SuppInfo. The gel electrophoresis figure of the RT-PCR products of the artificially inoculated samples
10337_2015_2931_MOESM3_ESM.docx (18 kb)
Supplementary material 3 (DOCX 17 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Jia Ruan
    • 1
  • Cheng Jun Sun
    • 1
    • 2
  • Fan Chen
    • 1
  • Chen Zhou
    • 1
  • Yuan Qian Li
    • 1
    • 2
  • Ya Pan Liu
    • 1
  • Yong Xin Li
    • 1
    • 2
  1. 1.West China School of Public HealthSichuan UniversityChengduChina
  2. 2.Sichuan Provincial Key Laboratory for Food Safety Monitoring and Risk AssessmentChengduChina

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