Full-Text Search:
Home|Journal Papers|About CNKI|User Service|FAQ|Contact Us|中文
《Biotechnology》 2005-03
Add to Favorite Get Latest Update

Two PCR Amplification Methods Influence the Membrane Chip Hybridizations Results for Detection of Group A Rotavirus

HUANG Hai-yan1,HAN Jin-xiang 1,WANG Jian-wei 2,3 , ZHU Bo 1 (1.Shandong Medical Biotechnology Center,Key Laboratory for Biotech-Drugs Ministry of Health,Jinan 250062; 2.Shandong Provincial Institute of Virology,Shandong Provincial Key Laboratory for Medical Virology Jinan 250062; 3.Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 100052,P.R.China)  
Group A rotavirus was the most important etiological agents of severe diarrhoeal illness of infants and young children worldwide.In this study,a reverse transcirptase-polymerase seminested chain reaction (RT-seminested PCR) using digoxigenin(dig)-labeled up-prime and oligoprobes distinctive to group A RV using chemiluminescence has been developed to detect group A RV.Two different PCR methods,seminested symmetry PCR and seminested asymmetry PCR,were adopted respectively.The results suggested both methods could produce the aimed size nucleotide sequences according to the agarose gel electrophoresis but different hybridization signals.The hybridization of seminested symmetry PCR product didnt show the ideal signals.While the asymmetry PCR could produce adequate single-strand nucleotide and display ideal hybridization signals , remarkably improved the detection sensitivity for group A RV. This study indicated that the seminested asymmetry PCR was a robust method to obtain abundant single-strain nucleotide for the use of chip hybridization,especially for the sequences contain of AT-rich.
【Fund】: “十五”国家重大科技专项项目资助(“食品安全关键技术研究” 2001BA804A22)
【CateGory Index】: R446.5
Download(CAJ format) Download(PDF format)
CAJViewer7.0 supports all the CNKI file formats; AdobeReader only supports the PDF format.
©2006 Tsinghua Tongfang Knowledge Network Technology Co., Ltd.(Beijing)(TTKN) All rights reserved