以囊胚培養液偵測囊胚DNA仍不可靠  最大問題為母體DNA之汙染干擾胚胎針測之準確性

Fertil Steril. 2018 Oct;110(5):870-879.e5. doi: 10.1016/j.fertnstert.2018.05.031.

Diagnostic efficacy of blastocoel fluid and spent media as sources of DNA for preimplantation genetic testing in standard clinical conditions.

Capalbo A1, Romanelli V2, Patassini C2, Poli M3, Girardi L2, Giancani A4, Stoppa M4, Cimadomo D4, Ubaldi FM4, Rienzi L4.

Author information

Erratum in

• Erratum. [Fertil Steril. 2019]

Abstract

OBJECTIVE:

To determine whether blastocoel fluid (BF) or spent blastocyst medium (SBM) is a suitable template for genotype and/or karyotype assessment of in vitro fertilization-generated embryos.

DESIGN:

Prospective blinded study.

SETTING:

Genetic laboratory.

PATIENT(S):

From 26 patients undergoing preimplantation genetic testing (PGT) treatments, 103 trophectoderms (TE), 92 BF samples, and 72 SBM samples.

INTERVENTION(S):

The BF and SBM were retrieved at the time of TE biopsy. Two DNA extraction strategies were evaluated on independent BF and SBM samples. Further enrolled samples were processed using next-generation sequencing and quantitative polymerase chain reaction for assessment of monogenic disorders (PGT-M) or aneuploidy (PGT-A).

MAIN OUTCOME MEASURE(S):

DNA amplification and concordance rates across BF, SBM, and TE to assess diagnostic efficiency.

RESULT(S):

No differences were detected among the DNA extraction methods tested. In PGT-M tests, for BF and SBM, 2.9% and 20.8% of all samples, respectively, produced a diagnosis concordant with the corresponding TE (n = 2 of 69 and 15 of 72, respectively). The SBM samples were associated with higher discordance rates and higher artifacts/contamination detection compared with BF. In multiple occasions, the maternal mutated variant was detected in the SBM of homozygous wild-type embryos, showing evidence of maternal DNA persistence in culture medium. In PGT-A tests, BF analysis showed high amplification failure rates (65.2%) and an overall concordance rate of 37.5% among amplified samples.

CONCLUSION(S):

Based on current methodologies, BF and SBM genetic analyses do not provide sufficiently reliable results to be employed clinically. Until the risk of maternal contamination can be properly prevented, SBM should not be used for PGT-M purposes.