Method and compositions for detecting epidermal growth factor receptor variant forms in cancer cells

摘要

Method and compositions for screening for the presence of Epidermal Growth Factor Receptor variant 3 (EGFR(v3)) in a sample are described. The method comprises obtaining a sample containing a plurality of cells; hybridizing a set of chromosomal probes to the sample, wherein the set comprises an EGFR(v3)-probe and a probe to chromosome 7 different from an EGFR(v3)-probe; and visualizing the hybridization pattern of the set of chromosomal probes in the plurality of cells of the sample, wherein the presence of at least one copy of chromosome 7 lacking a hybridization signal of the EGFR(v3)-probe in at least one cell is indicative of the presence of the EGFR(v3) in the sample. The method and compositions are suitable for diagnosing the therapeutic outcome for treating a patient having a cancer with an anti-EGFR therapeutic agent and for screening a sample for a predisposition for forming an EGFR-associated cancer.

主权项

1. A method of screening for the presence of EGFR(v3) in a sample, the method comprising:
(a) obtaining a sample containing a plurality of cells; (b) hybridizing a set of chromosomal probes to the sample, wherein the set comprises an EGFR (v3)-probe and a probe to chromosome 7 different from an EGFR(v3)-probe; and (c) visualizing a hybridization pattern of the set of chromosomal probes in the plurality of cells of the sample, wherein the hybridization pattern comprises a presence of a signal from the probe to chromosome 7 different from an EGFR(v3)-probe and an absence of a signal from the EGFR(v3)-probe in at least one cell, wherein the EGFR(v3)-probe is labeled amplicon 4 prepared according to the following procedure:
(i) contacting a DNA comprising a sequence encoding a wild-type EGFR gene with isolated oligonucleotide primers corresponding to SEQ ID NO:7 and SEQ ID NO:8;(ii) amplifying a portion of the sequence encoding a wild-type EGFR gene with isolated oligonucleotide primers corresponding to SEQ ID NO:7 and SEQ ID NO:8 in a polymerase chain reaction to form amplicon 4; and(iii) incorporating at least one label into amplicon 4 to form labeled amplicon 4.