Materials and methods for the diagnosis of pediatric tumors
OwnerDavid F. Carpentieriinventor
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Abstract
The present invention is directed to methods wherein the cytolocalization of WT1 protein can be used as a tool in the differential diagnosis of soft tissue tumors. Specifically, the invention is directed to the novel finding that elevated levels of WT1 protein in the cytoplasm of cells derived from a soft tissue tumor sample provide a positive diagnostic indicator for rhabdomyosarcoma (RMS).
Claims39
A method for diagnosing pediatric soft tissue tumors based on differential localization of a WT1-associated molecule, said method comprising:
- a) obtaining a biological sample from a pediatric patient;
- b) contacting said sample with an agent having affinity for said WT1-associated molecule and;
- c) determining a cellular localization of said WT1-associated molecule as indicated by localization of said agent having affinity for said WT1-associated molecule, wherein detection of elevated cytoplasmic levels of said WT1-associated molecule provides a positive diagnostic indicator of a rhabdomyosarcoma tumor.
A method for diagnosing pediatric soft tissue tumors based on differential localization of WT1 protein, said method comprising:
- a) obtaining a sample from a pediatric patient;
- b) contacting said sample with an antibody or antibody fragment immunologically specific for WT1 protein;
- and
- c) determining cellular localization of WT1 as indicated by localization of said antibody or antibody fragment immunologically specific for WT1, wherein elevated cytoplasmic WT1 staining provides a positive diagnostic indicator of a rhabdomyosarcoma tumor.
A method for detecting WT1 encoding nucleic acid in a biological sample as a tumor marker for rhabdomyosarcoma cancer, wherein said biological sample is derived from a patient diagnosed with rhabdomyosarcoma, said method comprising:
- a) extracting nucleic acids from said biological sample;
- b) contacting said extracted nucleic acid with oligonucleotide primers which specifically hybridize to WT1 encoding nucleic acids if any are present;
- c) subjecting said extracted nucleic acid and primers to conditions suitable for polymerase chain reaction amplification;
- and
- d) assessing the resulting reaction product for amplified WT1 nucleic acid.
A kit for detecting WT1 in a biological sample, said kit comprising:
- a) an antibody or fragment thereof immunologically specific for a region of WT1;
- b) a detectable label for said antibody;
- and
- c) reagents suitable for detecting WT1-antibody immunocomplexes, if present in said biological sample.
A kit for identifying WT1 in a biological sample, said kit comprising:
- a) at least one pair of primers, said primers having the sequence selected from the group consisting of:
- SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, and SEQ ID NO:
- 4;
- b) a polymerase enzyme suitable for use in polymerase chain reaction;
- c) buffers and nucleotides suitable for performing amplification reactions;
- d) a DNA sample comprising a positive control;
- and
- e) optionally an instruction protocol.
Dependencies are inferred from the claim text.