FOOD SCIENCE ›› 2009, Vol. 30 ›› Issue (20): 336-339.doi: 10.7506/spkx1002-6300-200920074

Previous Articles     Next Articles

Fluorescent RT-PCR Assay for Detection of Sapovirus in Shellfish

DENG Tian-ning1,PAN Liang-wen2,*,LU Rong2,GAO Qin2,ZHANG Shu-ya2,LU Zhong-shan2   

  1. 1. School of Bioengineering, East China University of Science and Technology, Shanghai 200237, China ;
    2. Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai 200135, China
  • Received:2009-01-15 Revised:2009-07-06 Online:2009-10-15 Published:2010-12-29
  • Contact: PAN Liang-wen E-mail:panlw888@126.com

Abstract:

In order to detect GI, GII, GIV and GV sapoviruses in shellfish, a fluorescent real-time polymerase chain reaction (RT-PCR) asssy was developed. sapoviruses in shellfish were concentrated using polyethylene glycol 8000 (PEG 8000) first. Total RNA of sapoviruses from Shellfish were extracted with Tri-reagent and purified with Poly(dT)25 magnetic beads due to its 3’ Poly(A) tails. Four genotypes of RNA with high purity were determined using the fluorescent RT-PCR assay. This method is effective and sensitive to obtain 101 copies as the limit of detection. Therefore, this fluorescent RT-PCR can be used in daily detection.

Key words: sapovirus, RT-PCR, detection

CLC Number: