乙脑病毒rt - ingenta connect.pdfVIP

! 1928 ! Ch i J Pharm A al 2010, 30( 10) * RT- PCR ** 高正琴, 岳秉飞, 贺争鸣 (, 100050) : RT- PCR : , C 1 , RNA, cDNA, PCR , , , 90 : RT- PCR, C DNA, 10 pg DNA90 RT- PCR, 23 : PCR, : ; ; ; ; : R917 : A : 0254- 1793( 2010) 10- 1928- 04 Developmentand application ofRT- PCR assay * ofJap anese encep ha litis virus * * GAO Zhe g- qi, YUE Bi g- fei, HE Zhe g- m i g ( Natio al I stitute for the Co trol of Pharm aceutical a d Biological Products, Beiji g 100050, China) Abstract Objective: To establish a RT - PCR assay for detectio of Japanese encephalitis virus( JEV ). Meh tods:A pairs of prmi ers specific to capsid protei C ge e of JEV SA 14 w ere desig ed accord i g to the published JEV complete ge ome seque ce, w ithw hich theRNA of JEV w as extracted a d reversely tra scribed to cDNA as a temp late for PCR amplificatio . Results:The established RT - PCR assay w as optmi ized, verified for specificity a d se sitivity, a d itsm i mi um lmi it usi g the recombi atio p lasm id DNA co tai i g JEV capsid protei C ge e as temp late w as 10 pg DNA. N i ety cli ical samp lesw ere detected by the established RT - PCR assay, from thirty- three of wh ich the specific target ge esw ere amplified. JEV w ere isolated a d ide tified from these brai tissues of m i i- pigs. Conclusion: The established RT - PCR assay laid a fou datio of detectio a d epidem iological i vesti gatio of JEV i fectio . ey words:Japanese encephalitis virus( JEV ); RT- PCR; specificity; se sitivity; detect ( Japa ese e cep