[ Objective] To improve the accuracy and efficiency of the detection which used in Perkinsus sp and Marteilia refringens, and then short- en the detective cycle. [ Method] According to the gene sequence of Perkinsus s...[ Objective] To improve the accuracy and efficiency of the detection which used in Perkinsus sp and Marteilia refringens, and then short- en the detective cycle. [ Method] According to the gene sequence of Perkinsus sp and Marteilia refringens from gene bank, design two pairs of spe- cific primers and two TaqMan probes with different fluorophores labeled. Optimizing the reactive conditions and reagent concentration in order that establishing the duplex real-time PCR method for detecting Perkinsus sp and Marteilia refringens simultaneously. [ Result ] The sensitivity of the du- plex real-time PCR method which about Pertdnsus sp and Marteilia refringens is 40 template copies. After combine the templates of Perkinsus sp and Marteilia refringens with different concentrations, this method still could be detect this two protozoan efficiently and synchronously. [ Condudon] The es- tablished duplex real-time PCR method for detecting Perkinsus sp. and Marteilia refringens possesses lots of advantages, such as specific, sensitive, rapid, quantitative and reproducible, can be used for clinical detection of infection which was caused by Perkinsus sp. and Marteilia refringens.展开更多
The paper aimed to establish a duplex PCR method for simultaneous detection of Pseudomonas savastanoi pv. Phaseolicola （Psp） and Curtobaeterium /accumfadens pv. Flaccumfaciens （Cff）. Based on the argK gene of Psp ...The paper aimed to establish a duplex PCR method for simultaneous detection of Pseudomonas savastanoi pv. Phaseolicola （Psp） and Curtobaeterium /accumfadens pv. Flaccumfaciens （Cff）. Based on the argK gene of Psp in GenBank, the primers PSPF1/PSPR2 were designed. The duplex PCR assay was dereloped using the combined primers PSPF1/PSPR2 and CflF1/CffR2, which were specific primers for Cff. The reaction conditions were optimized and specificity md sensitivity of the duplex PCR were tested. The expected DNA fragment was specifically amplified from the genomic DNA of Psp and Cff. Specificity was conirmed in the artificially inoculated soybean samples imparted. Thus, the duplex PCR developed in this study could be used for the simultaneous detection of Psp md Cff from imported soybean.展开更多
文摘[ Objective] To improve the accuracy and efficiency of the detection which used in Perkinsus sp and Marteilia refringens, and then short- en the detective cycle. [ Method] According to the gene sequence of Perkinsus sp and Marteilia refringens from gene bank, design two pairs of spe- cific primers and two TaqMan probes with different fluorophores labeled. Optimizing the reactive conditions and reagent concentration in order that establishing the duplex real-time PCR method for detecting Perkinsus sp and Marteilia refringens simultaneously. [ Result ] The sensitivity of the du- plex real-time PCR method which about Pertdnsus sp and Marteilia refringens is 40 template copies. After combine the templates of Perkinsus sp and Marteilia refringens with different concentrations, this method still could be detect this two protozoan efficiently and synchronously. [ Condudon] The es- tablished duplex real-time PCR method for detecting Perkinsus sp. and Marteilia refringens possesses lots of advantages, such as specific, sensitive, rapid, quantitative and reproducible, can be used for clinical detection of infection which was caused by Perkinsus sp. and Marteilia refringens.
基金Supported by Science and Technology Project of AQSIQ(2013IK277)National Rice Industry System Development Program
文摘The paper aimed to establish a duplex PCR method for simultaneous detection of Pseudomonas savastanoi pv. Phaseolicola （Psp） and Curtobaeterium /accumfadens pv. Flaccumfaciens （Cff）. Based on the argK gene of Psp in GenBank, the primers PSPF1/PSPR2 were designed. The duplex PCR assay was dereloped using the combined primers PSPF1/PSPR2 and CflF1/CffR2, which were specific primers for Cff. The reaction conditions were optimized and specificity md sensitivity of the duplex PCR were tested. The expected DNA fragment was specifically amplified from the genomic DNA of Psp and Cff. Specificity was conirmed in the artificially inoculated soybean samples imparted. Thus, the duplex PCR developed in this study could be used for the simultaneous detection of Psp md Cff from imported soybean.
