Abstract 2091: Ultrasensitive detection of mutations in the androgen receptor gene by ICE COLD-PCR
Cancer Research(2014)
摘要
Proceedings: AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012; Chicago, IL
The Androgen Receptor plays an important role in prostate cancer, androgen-dependent (AD) and androgen-independent (AI) disease progression. Anti-androgen therapies are the hallmark of prostate cancer treatment; however spontaneous AR mutations are often detected in hormone-refractory, androgen-ablated and metastatic tumors. ICE COLD-PCR (Improved & Complete Enrichment CO-amplification at Lower Denaturation temperature) is a technology that preferentially enriches mutant DNA sequences in an excess of wild-type DNA through selective amplification of the mutant DNA population using an oligonucleotide complementary to wild-type sequence (RS-oligo). This RS-oligo prevents PCR amplification of wild-type sequences while allowing amplification of DNA containing any mutation covered in the RS-oligo region. We have developed an ICE COLD-PCR assay for the enrichment of mutations in the Androgen Receptor including the AR H875R, H875Y and T878A mutations. The mutant-enriched DNA was then analyzed by standard Sanger DNA sequencing. The analytical sensitivity for the ICE COLD-PCR assay was 1 in 10,000 (0.01%) for all the mutations, compared to 20% limit of detection by standard Sanger sequencing. The ICE COLD-PCR assay developed here provides a means to detect low level mutations in the Androgen Receptor and thus will provide a valuable methodology for the early diagnosis, treatment guidance, outcome prediction and relapse monitoring of prostate cancer.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2091. doi:1538-7445.AM2012-2091
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关键词
androgen receptor gene,mutations,ultrasensitive detection,cold-pcr
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