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《Chinese Journal of Preventive Veterinary Medicine》 2008-12
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Development and application of Taq Man-MGB fluorescence quantitative RT-PCR assay for detection of porcine reproductive and respiratory syndrome virus

WEI Tian-chao1,3,TIAN Zhi-jun1,AN Tong-qing1,ZHOU Yan-jun1,XIAO Yan1,JIANG Yi-feng1,HAO Xiao-fang1,ZHANG Shan-rui1,PENG Jin-mei1,QIU Hua-ji1,TONG Guang-zhi1,2*(1.Division of Swine Infectious Diseases,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences,Harbin 150001,China;2.Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai 200232,China;3.College of Animal Science and Technology,Guangxi University,Nanning 530005,China)  
A quantitative Taq Man reverse transcription-polymerase chain reaction(RT-PCR) for detecting PRRSV was deve-loped using specific primers and fluorogenetic probe based on the ORF7 gene of porcine reproductive and respiratory syndrome virus(PRRSV).The established method was highly specific,sensitive and reproducible.It had a linear dynamic range of the 101 copies/μL to 107 copies/μL and could detect the least 10 copies of target DNA and 1 TCID50 virus.The assay was succesffuly used to test on serum and tissue samples collected from experimentally infected pigs at various times following infection.Pigs infected with the fifth passage PRRSV exhibitedd a rapid increase in virus load in serum on day 3 and reached a peak of 1010 copies/mL on day 7,while pigs infected with the sixty-fifth passage virus had a lower virus load which peaked at 106 copies/mL on day 21.The virus concentrations in the lung were 109 copies/g,which was much higher than that in other tissues.
【Fund】: 国家“973”计划(2005CBZ523200);; 十一五国家科技支撑计划项目(2006BAD06A03/01/04 2007BAD86B03)资助;; 国家自然科学基金(30470072)
【CateGory Index】: S852.65
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