Development and validation of an accurate quantitative real-time polymerase chain reaction–based assay for human blastocyst comprehensive chromosomal aneuploidy screening  

Oleh:

Treff, Nathan R. ; Tao, Xin ; Ferry, Kathleen M. ; Su, Jing

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Fertility and Sterility (keterangan: ada di ClinicalKey) vol. 97 no. 04 (Apr. 2012), page 819-824.
Topik: Quantitative PCR; Real-Time PCR; Aneuploidy Screening; Comprehensive Chromosome Screening

Ketersediaan

Perpustakaan FK Nomor Panggil: F02.K.2012.01 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

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Isi artikel

Objective To develop and validate a quantitative real-time polymerase chain reaction (qPCR)–based method for blastocyst trophectoderm comprehensive chromosome screening (CCS) of aneuploidy. Design Prospective, randomized, and blinded. Setting Academic center for reproductive medicine. Patient(s) Nine cell lines were obtained from a commercial cell line repository, and 71 discarded human blastocysts were obtained from 24 IVF patients that underwent preimplantation genetic screening. Intervention(s) None. Main Outcome Measure(s) Consistency of qPCR diagnosis of aneuploidy compared with either conventional karyotyping of cell lines or microarray-based diagnoses of human blastocysts. Result(s) Samples from nine cell lines with well characterized karyotypes were diagnosed by qPCR with 97.6% (41/42) consistency. After applying a minimum threshold for concurrence, 100% consistency was achieved. Developmentally normal blastocysts designated as aneuploid or arrested blastocysts designated as euploid by single-nucleotide polymorphism microarray analyses were assigned identical 24 chromosome diagnoses by qPCR in 98.6% of cases (70/71). Overall euploidy (n = 37) and aneuploidy (n = 34) were assigned with 100% consistency. Data was obtained for both sample types in 4 hours. Conclusion(s) These data demonstrate the first qPCR technology capable of accurate aneuploidy screening of all 24 chromosomes in 4 hours. This methodology provides an opportunity to evaluate trophectoderm biopsies with subsequent fresh euploid blastocyst transfer. Randomized controlled trials to investigate the clinical efficacy of qPCR-based CCS are currently underway.

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