摘要:
The present invention provides a method of identifying origin of a metastasis of unknown origin by obtaining a sample containing metastatic cells; measuring Biomarkers associated with at least two different carcinomas; combining the data from the Biomarkers into an algorithm where the algorithm normalizes the Biomarkers against a reference; and imposes a cut-off which optimizes sensitivity and specificity of each Biomarker, weights the prevalence of the carcinomas and selects a tissue of origin determining origin based on highest probability determined by the algorithm or determining that the carcinoma is not derived from a particular set of carcinomas; and optionally measuring Biomarkers specific for one or more additional different carcinoma, and repeating the steps for additional Biomarkers.
摘要:
A method for detecting the presence of a target sequence comprising: a. adding a probe, said probe having been produced by a method comprising: i. obtaining double strand polynucleotides known to contain complementary target sequences and repetitive sequences; ii. fragmenting said double strand polynucleotides into fragments; iii. denaturing said fragments into single strands; iv. hybridizing said repetitive sequences to form a mixture of double strands and single strands; v. cleaving said double strands; and vi. amplifying said single strands wherein said single strands are complementary to said target sequences; and
b. detecting said probe wherein said detection is selected from a group consisting of ISH, FISH, CGH, spectral karyotyping, chromosome painting, Northern blots, Southern blots, microarray analysis, and combinations thereof.
摘要:
The disclosed edge-blocker oligonucleotide based AS-NEPB-PCR method amplifies allele specific DNA (or RNA) while dramatically blocking amplification of wild type (WT) DNA (or RNA). The AS-NEPB-PCR design allows ready modification of an existing PCR reaction setup with an edge-blocker oligonucleotide together with an allele specific primer complementary to the mutant sequence to achieve allele specific amplification. The method simplifies assay optimization procedures and achieved sensitivity sufficient to detect a signal present at 0.1% level with close to 100% specificity, which is useful in detecting SNP or genetic mutations. The method was used to detect three different genetic mutations in cancer, in KRAS, BRAF, and EGFR, with three different types of modified edge-blocker oligonucleotides (phosphate, inverted dT and amino-C7). It was possible to detect one copy of mutant DNA in 1000-copy of normal DNA background of a heterogeneous sample, and was far more sensitive than the other blocking method.
摘要:
The invention includes methods for isolating circulating multiple myeloma cells as well as method of treating patients suspected of having diseases of abnormal plasma cells.