Methods for detection and quantification of EGFRvIII in the peripheral blood of GBM patients

A kit comprising:

• a first primer set comprising forward primers corresponding to a DNA sequence in intron 1 of an EGFR gene and a reverse primer that corresponds to a DNA sequence in exon 8 of an EGFR gene;
• a second primer set comprising forward primers corresponding to the DNA sequence in intron 1 of the EGFR gene that are separated by about 5 kb from the forward primers of the first primer set and the reverse primer that corresponds to a DNA sequence in exon 8 of the EGFR gene, wherein the forward primers of the first and second sets are comprised within the base pairs defining intron 1 of the EGFR gene;
• one or more amplification primers that hybridize to priming sites that flank breakpoints in EGFRvIII, wherein the one or more amplification primers are designed to yield a PCR fragment of about 300 base pairs and wherein the one or more amplification primers comprise SEQ ID NO:
• 29, SEQ ID NO:
• 30, SEQ ID NO:
• 31, or SEQ ID NO:
• 32;
• and
• a dual-labeled hydrolysis probe specific to a region of EGFR sequence amplified by the plurality of forward primers and the reverse primer.