Patent ID: 11913079
Assignee: QUEST DIAGNOSTICS INVESTMENTS INCORPORATED
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 18:
19. A method for detecting a dysregulation in an RET gene in a test sample, comprising:
(a) amplifying portions of a 5′ region of a transcript of the RET gene or a cDNA derived therefrom, if present in the test sample, with two or more different 5′ target primer pairs selected from:
a forward primer of SEQ ID NO: 41 and a reverse primer of SEQ IN NO: 42;
a forward primer of SEQ ID NO: 43 and a reverse primer of SEQ IN NO: 44;
a forward primer of SEQ ID NO: 45 and a reverse primer of SEQ IN NO: 46;
a forward primer of SEQ ID NO: 47 and a reverse primer of SEQ IN NO: 48; and
a forward primer of SEQ ID NO: 49 and a reverse primer of SEQ IN NO: 50;
(b) amplifying portions of a 3′ region of the transcript of the RET gene or a cDNA derived therefrom, if present in the test sample, with two or more different 3′ target primer pairs that are directed to the portions of the 3′ region of the RET gene,
(c) detecting the amplification products produced by the two or more 5′ target primer pairs and the two or more 3′ target primer pairs,
(d) determining the average cycle threshold (Ct) among the two or more 5′ target primer pairs and the average Ct among the two or more 3′ target primer pairs,
(e) calculating an IDE Score as the difference between the average cycle threshold among the 5′ target primer pairs and the average cycle threshold among the 3′ target primer pairs, and
(f) identifying the test sample as having a target gene dysregulation if the IDE Score is significantly different than a cutoff value and the difference indicates the presence of a RET gene dysregulation.