ESHRE PGD consortium best practice guidelines for amplification-based PGD

Abstract

In 2005, the European Society for Human Reproduction and Embryology (ESHRE) PGD Consortium published a set of Guidelines for Best Practice PGD to give information, support and guidance to potential, existing and fledgling PGD programmes. The subsequent years have seen the introduction of a number of new technologies as well as the evolution of current techniques. Additionally, in light of recent advice from ESHRE on how practice guidelines should be written and formulated, the Consortium believed it was timely to revise and update the PGD guidelines. Rather than one document that covers all of PGD, as in the original publication, these guidelines are separated into four new documents that apply to different aspects of a PGD programme, i.e. Organization of a PGD centre, fluorescence in situ hybridization-based testing, Amplification-based testing and Polar Body and Embryo Biopsy for PGD/preimplantation genetic screening. Here, we have updated the sections that pertain to amplification-based PGD. Topics covered in this guideline include inclusion/exclusion criteria for amplification-based PGD testing, preclinical validation of tests, amplification-based testing methods, tubing of cells for analysis, set-up of local IVF centre and Transport PGD centres, quality control/quality assurance and diagnostic confirmation of untransferred embryos.

PGD for hereditary breast and ovarian cancer: the route to universal tests for BRCA1 and BRCA2 mutation carriers

“…The robustness of the multiplex BRCA PCR tests are in accordance with the latest edition of the ESHRE PGD consortium guidelines, 16 with a PCR efficiency 490% and ADO rates o10%.…”

“…All primers were mixed in one multiplex PCR following the ESHRE guidelines. 16 After optimization of the single-cell multiplex PCR, all protocols were validated by testing at least 50 single leucocytes heterozygous for each marker in at least three separate experiments to assess amplification efficiency and ADO rate. Additionally 15-20 PCR blanks were analyzed to determine contamination.…”

The experience of 3 years of external quality assessment of preimplantation genetic diagnosis for cystic fibrosis

“…[2][3][4] With respect to monogenic diseases, PGD can theoretically be applied for any genetic disease with a definitive molecular diagnosis and/or defined linkage within a family. The quantity of sample available for the genetic analysis in PGD is minimal, and in most cases comprises no more than a single cell.…”

“…All technical aspects of the method should be addressed before the clinical application of any PGD PCR protocol, to ensure a reliable and accurate genotype result. 4 Besides the PCR-related pitfalls, adverse outcomes subsequent to a PGD diagnosis have been attributed to tube switching during the genotype analysis, transfer of the wrong embryo(s), inappropriate protocol design and chromosomal mosaicism in the embryo(s) being analysed. 5 During the two decades that PGD has been applied as a clinical diagnostic service, the PGD community has accumulated considerable experience, which indicates that the potential causes of misdiagnosis can be substantially addressed through careful assay design of optimised fluorescent and multiplexed robust PCR protocols, overall applied with the most stringent laboratory procedures to additionally preclude contamination and tube-switching.…”

“…5 During the two decades that PGD has been applied as a clinical diagnostic service, the PGD community has accumulated considerable experience, which indicates that the potential causes of misdiagnosis can be substantially addressed through careful assay design of optimised fluorescent and multiplexed robust PCR protocols, overall applied with the most stringent laboratory procedures to additionally preclude contamination and tube-switching. 4,5 However, the absence of universally standardsed protocols, compounded by the frequent need to set up case-specific protocols, especially for rare monogenic diseases, means that each PGD centre has its own 'in-house' protocols. This may present issues with quality control, standardisation of laboratory protocols and meeting accreditation standards ISO 15189:2007. www.iso.org/iso/home/store/catalogue_ics/ catalogue_detail_ics.htm?csnumber=42641.…”