Epigenomics Ag

New Day Diagnostics, LLC

Juergen Distler

Thomas Hildmann

Xiaoling Song

Reimo Tetzner

Dimo Dietrich

Matthias Schuster

Kurt Berlin

Matthias Ballhause

Andrew Sledziewski

Jennifer Maas

Cathy Lofton-day

Volker Liebenberg

Fabian Model

Andrew Z Sledziewski

Catherine E Lofton-Day

Ralf Lesche

Jörn Lewin

Anne Schlegel

Denise Kottwitz

Shannon Payne

Susan Cottrell

Jürgen Distler

Rene Cortese

Joern Lewin

Michael Krouse

Theo De Vos

Antje Kluth

Cathy E Lofton-Day

Theo Devos

Jesse Ho

Philipp Schatz

Joe Lograsso

Michael Wandell

Catherine Lofton-Day

Cathy Lofton-Day

The present invention relates to the field of pharmacogenomics and in particular to detecting the presence or absence of hypermethylated DNA. The detection of CpG methylation in marker DNA is useful for the diagnosis of cancers and the invention provides improved methods for this purpose. These improved methods allow in particular for a more sensitive detection of methylated marker DNA with high backgrounds of unmethylated marker DNA.

Methods for detecting CpG methylation and for diagnosing cancer

The invention provides methods, nucleic acids and kits for detecting, or for detecting and distinguishing between or among liver cell proliferative disorders or for detecting, or for detecting and distinguishing between or among colorectal cell proliferative disorders. The invention discloses genomic sequences the methylation patterns of which have utility for the improved detection of and differentiation between said class of disorders, thereby enabling the improved diagnosis and treatment of patients.

Methods and nucleic acids for analyses of cellular proliferative disorders

Aspects of the present invention relate to compositions and methods for providing DNA fragments from a remote sample. In particular aspects a remote sample comprising DNA is provided, DNA is isolated from the remote sample, and the isolated DNA is treated in a way which allows differentiation of methylated and unmethylated cytosine. Additional, particular embodiments provide compositions and methods for methylation analysis of DNA derived from a remote sample. Other aspects provide for compositions and methods of whole genome amplification of bisulfite treated DNA. Other aspects provide methods for determining the presence or absence of methylation of at least one cytosine, or a series of cytosines in cis, in human DNA of a blood sample, a plasma sample, a serum sample or a urine sample from a human individual.

Method for determining the presence or absence of methylation in a sample

The invention relates to a method for analyzing cytosine methylations in DNA sequences, according to which non-methylated cytosines are first converted into uracil while 5-methylcytosine remains unmodified. The DNA is then amplified by means of a polymerase and at least one primer whose 5 end is connected to a probe via a linker. The probe is intramolecularly hybridized onto the amplified products in accordance with the methylation state of the DNA, hybridization being detectable via different detection systems. The inventive method is particularly suitable for diagnosing and predicting cancer diseases and other diseases associated with a modification of the methylation state as well as for predicting undesired effects of medicaments.

Method for the detection of cytosine methylations in DNA

The invention provides methods, nucleic acids and kits for detecting prostate cell proliferative disorders. The invention discloses genomic sequences the methylation patterns of which have utility for the improved detection of said disorder, thereby enabling the improved diagnosis and treatment of patients.

Methods and nucleic acids for the analysis of gene expression associated with the development of prostate cell proliferative disorders

Methods and nucleic acids for the analyses of cellular proliferative disorders

Aspects of the invention provide methods, nucleic acids and kits for detecting, or for detecting and distinguishing between or among liver cell proliferative disorders or for detecting, or for detecting and distinguishing between or among colorectal cell proliferative disorders. Particular aspects disclose and provide genomic sequences the methylation patterns of which have substantial utility for the improved detection of and differentiation between said class of disorders, thereby enabling the improved diagnosis and treatment of patients.

The present invention relates to a method for methylation analysis. It comprises the providing of a double stranded nucleic acid; its conversion, whereby unmethylated bases become distinguishable in their base-pairing behavior from methylated bases, and the analysis of both of the converted nucleic acid strands.

Method for methylation analysis of nucleic acid

Particular aspects provide methods for specific amplification of template DNA in the presence of potentially contaminating PCR products from previous amplification experiments. Particular embodiments comprise, in a first step, contacting DNA with a bisulfite solution, which sulfonates unmethylated (but not methylated) cytosines, resulting in cytosine deamination and generation of sulfonated uracil. Such sulfonation protects the template nucleic acid from being a target for the enzyme uracil-DNA-glycosylase (UNG), whereas any contaminating DNA, which contains unprotected unsulfonated or desulfonated uracils, is degraded enzymatically while the UNG is active. After UNG treatment and inactivation thereof, the sulfonated uracil bases are converted into uracil by desulfonation. Such aspects have substantial utility for decontamination of nucleic acid samples; e.g., for avoiding amplification of 'carry over products' in the context of DNA methylation analysis. In further aspects, the inventive methods can be generally used as simplified methods of bisulfite treatment.

Method for the carry-over protection in DNA amplification systems targeting methylation analysis achieved by a modified pre-treatment of nucleic acids

The method according to the invention concerns in particular a method for the quantification of methylated DNA. For this purpose, the DNA to be examined is first transformed such that unmethylated cytosine is converted to uracil while 5-methylcytosine remains unchanged. Subsequently, the transformed DNA is amplified in the presence of a pair of real-time probes. For this, a probe is constructed, which is specific for the methylated or for the unmethylated state of the DNA, and a probe, which binds methylation-unspecifically to the amplificate. The ratio of the signal intensities of the probes or the CT values allows for the calculation of the degree of methylation of the examined DNA. The method according to the invention is suited particularly for the diagnosis and prognosis of cancer and other diseases associated with a change in the methylation status, as well as, prediction of adverse for side-effects of pharmaceuticals.

Method for the quantification of methylated DNA

Aspects of the invention relate to composition and methods for the providing of DNA for methylation analysis that is in particular suitable to be applied in reference laboratories. Further 5 aspects of the invention relate to composition and methods for the highly specific and sensitive methylation analysis of the Septin 9 gene also in particular suitable to be applied in reference laboratories.

Method for methylation analysis

Growing community of inventors

Berlin, Germany

Reimo Tetzner