[Objective] The aim was to explore the reasons of false positives in Different Display Reverse Transcription（DDRT）analysis.[Method] Soybean varieties ＂Jilin 30＂ and ＂Tongnong 13＂ were used as materials to carry ...[Objective] The aim was to explore the reasons of false positives in Different Display Reverse Transcription（DDRT）analysis.[Method] Soybean varieties ＂Jilin 30＂ and ＂Tongnong 13＂ were used as materials to carry out analysis on false positives in DDRT analysis.[Result] An important origin of false positives appeared in DDRT analysis was the non-specific amplification caused by the combination of single primer and cDNA.The parallel PCR test of single primer should be set so as to verify whether the obtained fragments were the false positives or the PCR productions combined with single primer.[Conclusion] This study had provided basis for improving the success rate of DDRT experiment.展开更多
YN87448 virus strain was isolated from a fevered female patient (52 years old ) in Yunnan Province in 1986, and was identified as a member of Alphavirus using serological method. One primer was designed from common ha...YN87448 virus strain was isolated from a fevered female patient (52 years old ) in Yunnan Province in 1986, and was identified as a member of Alphavirus using serological method. One primer was designed from common hairpin conserved region of Alphavirus. Two fragments were amplified by single primer differential RT PCR, and they were cloned into pGEM T vector. Sequences were determined, and then analyzed by Software and GenBank. Results showed that the 1118pb long fragment was highly homologous to the sequence of Sindbis like virus S.A.A.R86. The homogeneity of the 1118bp fragment between YN87448 virus strain and S.A.A.R86 virus strain was 98%. Single primer differential RT PCR is a useful method with simple, economic, and practical value for Alphavirus identification.展开更多
文摘[Objective] The aim was to explore the reasons of false positives in Different Display Reverse Transcription（DDRT）analysis.[Method] Soybean varieties ＂Jilin 30＂ and ＂Tongnong 13＂ were used as materials to carry out analysis on false positives in DDRT analysis.[Result] An important origin of false positives appeared in DDRT analysis was the non-specific amplification caused by the combination of single primer and cDNA.The parallel PCR test of single primer should be set so as to verify whether the obtained fragments were the false positives or the PCR productions combined with single primer.[Conclusion] This study had provided basis for improving the success rate of DDRT experiment.
文摘YN87448 virus strain was isolated from a fevered female patient (52 years old ) in Yunnan Province in 1986, and was identified as a member of Alphavirus using serological method. One primer was designed from common hairpin conserved region of Alphavirus. Two fragments were amplified by single primer differential RT PCR, and they were cloned into pGEM T vector. Sequences were determined, and then analyzed by Software and GenBank. Results showed that the 1118pb long fragment was highly homologous to the sequence of Sindbis like virus S.A.A.R86. The homogeneity of the 1118bp fragment between YN87448 virus strain and S.A.A.R86 virus strain was 98%. Single primer differential RT PCR is a useful method with simple, economic, and practical value for Alphavirus identification.
