Vol.3 No.2 2010

66/86

Research paper : Development of an accurate and cost-effective quantitative detection method for specific gene sequences (N. Noda)−169−Synthesiology - English edition Vol.3 No.2 (2010) YesYesNoNoNoNoResistance to inhibitorsNot necessaryNot necessaryNecessaryNecessaryNecessaryNecessaryReal time PCR devicePossibleImpossiblePossiblePossiblePossibleImpossibleCheck amplified product by melting curve analysisNot necessaryNecessaryNot necessaryNot necessaryNot necessaryNot necessaryElectrophoresisNecessaryNecessaryNot necessaryNot necessaryNot necessaryNot necessaryInternal standard geneNeeded for eachtarget gene(label with 2 colors)Not necessaryCan deal with all targetgenes with one fluorescentprobe (label with 1 color)Not necessaryNeeded for eachtarget gene(label with 1 color)Needed for eachtarget gene(label with 2 colors)Fluorescent probe ABC methodCompetitive methodUniversal QProbemethodIntercalator methodQProbe methodTaqMan probemethodInternal standard methodReal time methodTable 1 Comparison of the characteristics of quantitative PCR methodsTable 2 Comparison of the universal QProbe method and the conventional RT-PCR method*Based on estimates by J-Bio 21 Corporation.○×○Multicolor detection○×○SNP genotyping○(Minimum next day)○(Minimum next day)×(1~2 weeks)Time needed for preparation*○(1 gene: about 6,000 yen)◎ (1 gene: about 2,000 yen)× (1 gene: 20,000 yen or higher)Cost* (probe, primer)○(Non specific product isnot detected)× (Non specific product isalso detected)○(Non specific product isnot detected)SpecificityIntercalator methodFluorescent probe methodUniversal QProbe methodConventional methodFig. 9 Scenario for the realization of the universal QProbe PCR and ABC-PCR methodsUniversal QProbe PCR methodABC-PCR methodAdvantage ofthe methodHigh specificity (advantage of probe method)Cost performance, high applicability, short preparation time of probe (advantage of joint DNA)Accuracy (resistant to amplification inhibitors)Non real time method (does not need RT-PCR thermal cycler)Points in realizationIntroduce as alternative to current RT-PCRPoints in realizationIntroduction of quantitative PCR at low costIntroduction of quantitative PCR with high accuracyCandidate product: Subcontract service for genetic analysis such as joint DNA, fluorescent probe, etc.Candidate products: Fluorescence measurement device, reagent kit, etc.Future prospectDevelopment of new technology combining universal QProbe and ABC methodsDevelopments of on-site and mobile gene quantification devices combining with the isothermal gene amplificationValue and impact on market ofthe candidate productShort preparation time needed for probe synthesisGenetic analysis subcontract service that enables quick data provision (quick delivery)Value and impact on market ofthe candidate productReduction in introduction cost of the gene quantification technology through marketing of inexpensive fluorescence measurement deviceAdvantage ofthe method

※このページを正しく表示するにはFlashPlayer9以上が必要です