Abstract: Provided are new and improved methods for detecting circulating tumor cells and tumor cell DNA in patient blood or other biofluid samples. Particular aspects comprise three steps: DNA extraction from patient samples, DNA digestion with multiple selected methylation-sensitive enzymes, and target amplification by a conventional or a real-time PCR with specific probe and/or primers. Also provided are a total of 40 tumor-specific DNA methylation loci as biomarkers having substantial utility and specificity in major types of human malignancies including hematopoietic and solid tumors.

Abstract: Differential Methylation Hybridization (DMH) was used to identify novel methylation markers and methylation profiles for hematopoieetic malignancies, leukemia, lymphomas, etc. (e.g., non-Hodgkin's lymphomas (NHL), small B-cell lymphomas (SBCL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma (B-CLL/SLL), chronic lymphocytic leukemia (CLL), multiple myeloma (MM), acute myelogenous leukemia (AML), acute lymphoblastic leukemia (ALL), etc.). Particular aspects provide novel biomarkers for NHL and subtypes thereof (e.g., MCL, B-CLL/SLL, FL, DLBCL, etc.), AML, ALL and MM, and further provide non-invasive tests (e.g. blood tests) for lymphomas and leukemias. Additional aspects provide markers for diagnosis, prognosis, monitoring responses to therapies, relapse, etc., and further provide targets and methods for therapeutic demethylating treatments.