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"Muu" projekt 037293
037293 "Quantitative Analysis of Genes in Single Cells (1.09.2006−31.08.2008)", Andres Metspalu, Eesti Biokeskus.
037293
037293
Quantitative Analysis of Genes in Single Cells
Quantitative Analysis of Genes in Single Cells
1.09.2006
31.08.2008
Teadus- ja arendusprojekt
Muu
ValdkondAlamvaldkondCERCS erialaFrascati Manual’i erialaProtsent
1. Bio- ja keskkonnateadused1.12. Bio- ja keskkonnateadustega seotud uuringud, näiteks biotehnoloogia, molekulaarbioloogia, rakubioloogia, biofüüsika, majandus- ja tehnoloogiauuringudT490 Biotehnoloogia 1.5. Bioteadused (bioloogia, botaanika, bakterioloogia, mikrobioloogia, zooloogia, entomoloogia, geneetika, biokeemia, biofüüsika jt100,0
AsutusRollPeriood
Eesti Biokeskuspartner01.09.2006−31.08.2008
AsutusRiikTüüp
European Commission
PerioodSumma
01.09.2006−31.08.20083 072 992,00 EEK (196 399,98 EUR)
196 399,98 EUR
0,00 EUR
FP6 LSHB-CT-2006

"The partners will develop methods and instruments to analyze the copy number of nucleic acid sequences down to the single cell / single molecule level, with the goal of developing an early diagnosis system for a child disease, namely hemophagocytic lymphohistiocytosis (HLH). The underlying technique is amplification based counting (ABC), enabling to quantify the copy number of genetic sequences with a resolution of about 100 base pairs in single cells. ABC is based on the discovery that there is a strong correlation between the number of positive PCRs from a genetic sequence and the sequence copy number if the sequence is present only in a few copies per sample. The method provides a resolution which is orders of magnitudes higher than for Fluorescence In Situ Hybridization and works quantitatively with much lower sample amounts than quantitative PCR. To prove ABC's effectiveness for clinical applications the partners will develop a single cell manipulation unit that picks cells from a solution and transfers them onto an integrated PCR and hybridization slide (AmpliGrid). The AmpliGrid contains dried-on PCR reagents as well as hybridization probes to detect the presence and specificity of the PCR products. The single cells on the AmpliGrid will then be processed automatically in an integrated PCR and hybridization machine (AmpliHyb). Clinical samples will be investigated in which copy number deviations are pathologic as in genetic diseases. As a model system QuAGSiC chose HLH which is hard to diagnose and fatal without specific therapeutic measures, as well as trisomy 21 which is relevant in prenatal and postnatal diagnostics. For these applications the project will cover the complete workflow from identifying the cells of interest to quantifying the copy number of nucleic acid sequences in clinical samples. Those diseases were selected as they will enable to demonstrate the efficiency of the proposed diagnostic assay as well as its broad range of applications."
"The partners will develop methods and instruments to analyze the copy number of nucleic acid sequences down to the single cell / single molecule level, with the goal of developing an early diagnosis system for a child disease, namely hemophagocytic lymphohistiocytosis (HLH). The underlying technique is amplification based counting (ABC), enabling to quantify the copy number of genetic sequences with a resolution of about 100 base pairs in single cells. ABC is based on the discovery that there is a strong correlation between the number of positive PCRs from a genetic sequence and the sequence copy number if the sequence is present only in a few copies per sample. The method provides a resolution which is orders of magnitudes higher than for Fluorescence In Situ Hybridization and works quantitatively with much lower sample amounts than quantitative PCR. To prove ABC's effectiveness for clinical applications the partners will develop a single cell manipulation unit that picks cells from a solution and transfers them onto an integrated PCR and hybridization slide (AmpliGrid). The AmpliGrid contains dried-on PCR reagents as well as hybridization probes to detect the presence and specificity of the PCR products. The single cells on the AmpliGrid will then be processed automatically in an integrated PCR and hybridization machine (AmpliHyb). Clinical samples will be investigated in which copy number deviations are pathologic as in genetic diseases. As a model system QuAGSiC chose HLH which is hard to diagnose and fatal without specific therapeutic measures, as well as trisomy 21 which is relevant in prenatal and postnatal diagnostics. For these applications the project will cover the complete workflow from identifying the cells of interest to quantifying the copy number of nucleic acid sequences in clinical samples. Those diseases were selected as they will enable to demonstrate the efficiency of the proposed diagnostic assay as well as its broad range of applications."
http://www.uni-ulm.de/quagsic/