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For the last 20 years, preimplantation genetic diagnosis (PGD) has been mostly performed on cleavage stage embryos after the
biopsy of 1–2 cells and PCR and FISH have been used for the diagnosis. The main indications have been single gene disorders
and inherited chromosome abnormalities. Preimplantation genetic screening (PGS) for aneuploidy is a technique that has used
PGD technology to examine chromosomes in embryos from couples undergoing IVF with the aim of helping select the chromosomally
‘best’ embryo for transfer. It has been applied to patients of advanced maternal age, repeated implantation failure, repeated
miscarriages and severe male factor infertility. Recent randomised controlled trials (RCTs) have shown that PGS performed
on cleavage stage embryos for a variety of indications does not improve delivery rates. At the cleavage stage, the cells biopsied
from the embryo are often not representative of the rest of the embryo due to chromosomal mosaicism. There has therefore been
a move towards blastocyst and polar body biopsy, depending on the indication and regulations in specific countries (in some
countries, biopsy of embryos is not allowed). Blastocyst biopsy has an added advantage as vitrification of blastocysts, even
post biopsy, has been shown to be a very successful method of cryopreserving embryos. However, mosaicism is also observed
in blastocysts. There have been dramatic changes in the method of diagnosing small numbers of cells for PGD. Both array-comparative
genomic hybridisation and single nucleotide polymorphism arrays have been introduced clinically for PGD and PGS. For PGD,
the use of SNP arrays brings with it ethical concerns as a large amount of genetic information will be available from each
embryo. For PGS, RCTs need to be conducted using both array-CGH and SNP arrays to determine if either will result in an increase
in delivery rates. 相似文献
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Jennifer Hodgetts Gaynor Johnson Kate Perkins Sioban Ostoja-Starzewska Neil Boonham Rick Mumford Matthew Dickinson 《Molecular biotechnology》2014,56(9):803-813
Partial recombinant secA proteins were produced from six different phytoplasma isolates representing five 16Sr groups and the expressed, purified recombinant (partial secA) protein from Cape St. Paul wilt disease phytoplasma (CSPWD, 16SrXXII) was used to immunise mice. Monoclonal antibodies (mAbs) were selected by screening hybridoma supernatants for binding to the recombinant proteins. To characterise the binding to proteins from different phytoplasmas, the antibodies were screened by ELISA and western blotting, and epitope mapping was undertaken. Eight different mAbs with varying degrees of specificity against recombinant proteins from different phytoplasma groups were selected. Western blotting revealed that the mAbs bind to proteins in infected plant material, two of which were specific for phytoplasmas. ELISA testing of infected material, however, gave negative results suggesting that either secA was not expressed at sufficiently high levels, or conformational changes of the reagents adversely affected detection. This work has shown that the phytoplasma secA gene is not a suitable antibody target for routine detection, but has illustrated proof of principle for the methodology. 相似文献
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Mammalian cells have developed complex mechanisms to identify DNA damage and activate the required response to maintain genome integrity. Those mechanisms include DNA damage detection, DNA repair, cell cycle arrest and apoptosis which operate together to protect the conceptus from DNA damage originating either in parental gametes or in the embryo's somatic cells. DNA repair in the newly fertilized preimplantation embryo is believed to rely entirely on the oocyte's machinery (mRNAs and proteins deposited and stored prior to ovulation). DNA repair genes have been shown to be expressed in the early stages of mammalian development. The survival of the embryo necessitates that the oocyte be sufficiently equipped with maternal stored products and that embryonic gene expression commences at the correct time. A Medline based literature search was performed using the keywords 'DNA repair' and 'embryo development' or 'gametogenesis' (publication dates between 1995 and 2006). Mammalian studies which investigated gene expression were selected. Further articles were acquired from the citations in the articles obtained from the preliminary Medline search. This paper reviews mammalian DNA repair from gametogenesis to preimplantation embryos to late gestational stages. 相似文献
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