Method for Detection of DNA Methyltransferase RNA in Plasma and Serum
AssigneeOncoMEDx, Inc.
InventorMichael S. Kopreski
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Abstract
The methods of the invention detect in a qualitative or quantitative fashion DNA methyltransferase RNA in blood plasma, serum, and other bodily fluids. The inventive methods are useful for aiding detection, diagnosis, monitoring, treatment, or evaluation of neoplastic disease, and for identifying individuals who have a predisposition to disease or who might benefit from further evaluation, monitoring or therapy.
Claims31
A method of detecting extracellular DNA methyltransferase 3b (DNMT3b) RNA in blood plasma or serum from a human or animal, the method comprising the steps of:
- a) extracting mammalian extracellular RNA from blood plasma or serum;
- b) amplifying or signal amplifying a portion of the extracted extracellular RNA or cDNA prepared therefrom, wherein said fraction comprises DNMT3b RNA, and wherein amplification is performed in either a qualitative or quantitative fashion using primers or probes specific for DNMT3b RNA or cDNA;
- and
- c) detecting the amplified DNMT3b RNA or cDNA product or signal.
A method of detecting DNA methyltransferase 3b (DNMT3b) RNA in blood or a blood fraction from a human or animal, the method comprising the steps of:
- a) extracting mammalian extracellular RNA from or a blood fraction from a human or animal;
- b) amplifying or signal amplifying a portion of the extracted extracellular RNA or cDNA prepared therefrom, wherein said fraction comprises DNMT3b RNA, and wherein amplification is performed in either a qualitative or quantitative fashion using primers or probes specific for DNMT3b RNA or cDNA;
- and
- c) detecting the amplified DNMT3b RNA or cDNA product or signal.
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A method of identifying a human or animal having DNA methyltransferase 3b (DNMT3b) RNA-expressing cells or tissue, the method comprising the steps of:
- a) extracting mammalian extracellular RNA from plasma or serum;
- b) amplifying or signal amplifying a portion of the extracted extracellular RNA or cDNA prepared therefrom, wherein said fraction comprises DNMT3b RNA, and wherein amplification is performed in a qualitative or quantitative fashion using primers or probes specific for DNMT3b RNA or cDNA;
- and
- c) detecting the amplified DNMT3b RNA or cDNA product or signal, wherein the presence thereof identifies a human or animal having DNA methyltransferase 3b (DNMT3b) RNA-expressing cells or tissue.
A method for detecting extracellular DNA methyltransferase 3b (DNMT3b) RNA, or cDNA reverse-transcribed therefrom, comprising the steps of extracting extracellular RNA comprising DNMT RNA from blood plasma or serum, with or without converting said extracellular RNA to cDNA, hybridizing said extracellular RNA or cDNA to a detectably-labeled probe specific for DNMT3b RNA or cDNA, and detecting hybridization of DNMT3b RNA or cDNA with the detectably-labeled probe.
A method for detecting extracellular DNA methyltransferase 3b (DNMT3b) RNA, or cDNA reverse-transcribed therefrom, comprising the steps of extracting extracellular RNA comprising DNMT RNA from a bodily fluid, with or without converting said extracellular RNA to cDNA, hybridizing said extracellular RNA or cDNA to a detectably-labeled probe specific for DNMT3b RNA or cDNA, and detecting hybridization of DNMT3b RNA or cDNA with the detectably-labeled probe.
A method for detecting a plurality of mammalian extracellular RNA species in plasma or serum from a human or animal, wherein one mammalian extracellular RNA species is a DNA methyltransferase 3b (DNMT3b) RNA species, the method comprising the steps of:
- a) extracting mammalian extracellular RNA from plasma or serum of a human or animal, wherein said extracted extracellular RNA comprises a plurality of mammalian RNA species, wherein one RNA species is DNMT RNA;
- b) amplifying or signal amplifying concurrently or sequentially at least one of said plurality of extracellular RNA or cDNA produced therefrom, wherein one of said extracellular RNA is DNMT3b RNA, to thereby produce an amplified product, wherein amplification is performed qualitatively or quantitatively using primers or probes specific for each RNA species;
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- c) detecting the amplified product produced from every amplified RNA species or cDNA produced therefrom.
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A diagnostic kit, comprising DNA methyltransferase (DNMT) RNA or cDNA specific amplification primers or probes.
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Dependencies are inferred from the claim text.