| 主权项 |
1. A method for monitoring the therapy of a patient suffering from glioblastoma multiforme (GBM), comprising:
(a) subjecting a sample from the patient comprising genomic DNA to long range polymerase chain reaction amplification of epithelial growth factor receptor vIII (EGFRvIII) gene, wherein the long range polymerase chain reaction amplification utilizes a plurality of forward primers corresponding to a DNA sequence in intron 1 of an epithelial growth factor receptor (EGFR) gene and a reverse primer that corresponds to a DNA sequence in exon 8 of an EGFR gene, such that if the sample comprises genomic DNA comprising EGFRvIII, a PCR product is formed, and if the sample does not comprise genomic DNA comprising EGFRvIII, a PCR product is not formed, wherein the plurality of forward primers are comprised within the base pairs defining intron 1 of the EGFR, each primer being separated by at least 5 kb from each other, and wherein the presence of PCR product is indicative of a presence of EGFRvIII in the sample, and the absence of PCR product is indicative of an absence of EGFRvIII in the sample; (b) identifying deletion breakpoints in the PCR product and designing amplification primers that hybridize to priming sites that flank the breakpoints, wherein the amplification primers are designed to yield a PCR fragment of about 300 base pairs; and (c) amplifying DNA from body fluid samples of said patient using the amplification primers to form an amplified DNA fragment of EGFRvIII. |
