PMID- 32590167
OWN - NLM
STAT- MEDLINE
DCOM- 20210510
LR  - 20210510
IS  - 1872-7654 (Electronic)
IS  - 0301-2115 (Linking)
VI  - 252
DP  - 2020 Sep
TI  - Noninvasive sexing of human preimplantation embryos using RT-PCR in the spent 
      culture media: A proof-of-concept study.
PG  - 89-93
LID - S0301-2115(20)30387-0 [pii]
LID - 10.1016/j.ejogrb.2020.06.023 [doi]
AB  - Preimplantation genetic testing (PGT) routinely requires biopsy which is an 
      invasive approach. The aim of this study was to examine a noninvasive approach 
      for sexing of preimplantation embryos using polymerase chain reaction 
      (PCR)/reverse transcriptase-PCR (RT-PCR) based on the presence of SRY DNA/RNA in 
      the spent culture medium. Two groups were evaluated: in group 1, 40 embryos of 
      routine PGT volunteers were cultured individually after biopsy and in group 2, 31 
      embryos were cultured individually until Day-5 post-fertilization. Each group was 
      further divided into three subgroups: RNA extraction (RE), nucleic acid (NA) and 
      DNase treated (DT). In the NA and DT subgroups, cDNA synthesis was performed 
      directly on culture medium with or without DNase treatment in DT and NA 
      subgroups, respectively. The results of sexing based on the PCR/RT-PCR in the 
      culture medium, were compared with the results of sexing by fluorescence in situ 
      hybridization (FISH) technique. In group 1, all samples were correctly diagnosed. 
      In group 2, five female samples were misdiagnosed. Test's sensitivity, 
      specificity and accuracy were 100 %, 94.44 % and 96.88 %, in RE, 100 %, 81.82 % 
      and 93.55 % in DT and 100 %, 71.43 % and 85.71 % in NA, respectively. 
      Preimplantation sexing without embryo biopsy, in the spent embryo culture media 
      using RNA amplification based methods including RE and DT seem to be more 
      reliable while nucleic acid based method, NA, led to the highest misdiagnoses 
      probably due to DNA contamination. Since all male samples were correctly 
      diagnosed in all subgroups of this preliminary study, preimplantation noninvasive 
      sexing on culture medium seems feasible, however all sources of nucleic acid 
      contamination must be carefully avoided.
CI  - Copyright (c) 2020 Elsevier B.V. All rights reserved.
FAU - Esmaeili, Mahnaz
AU  - Esmaeili M
AD  - Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute 
      for Reproductive Biomedicine, ACECR, Tehran, Iran; Department of Genetics, 
      Faculty of Basic Sciences and Advanced Technologies in Biology, Department of 
      Genetics, University of Science and Culture, ACECR, Tehran, Iran.
FAU - Bazrgar, Masood
AU  - Bazrgar M
AD  - Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute 
      for Reproductive Biomedicine, ACECR, Tehran, Iran. Electronic address: 
      mbazrgar@royaninstitute.org.
FAU - Gourabi, Hamid
AU  - Gourabi H
AD  - Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute 
      for Reproductive Biomedicine, ACECR, Tehran, Iran.
FAU - Ebrahimi, Bita
AU  - Ebrahimi B
AD  - Department of Embryology, Reproductive Biomedicine Research Center, Royan 
      Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
FAU - Boroujeni, Parnaz Borjian
AU  - Boroujeni PB
AD  - Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute 
      for Reproductive Biomedicine, ACECR, Tehran, Iran.
FAU - Fakhri, Mostafa
AU  - Fakhri M
AD  - Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute 
      for Reproductive Biomedicine, ACECR, Tehran, Iran.
LA  - eng
PT  - Journal Article
DEP - 20200612
PL  - Ireland
TA  - Eur J Obstet Gynecol Reprod Biol
JT  - European journal of obstetrics, gynecology, and reproductive biology
JID - 0375672
RN  - 0 (Culture Media)
SB  - IM
MH  - *Blastocyst
MH  - Culture Media
MH  - Female
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Male
MH  - Polymerase Chain Reaction
MH  - Pregnancy
MH  - *Reverse Transcriptase Polymerase Chain Reaction
MH  - *Sex Determination Analysis
OTO - NOTNLM
OT  - Culture medium
OT  - Noninvasive
OT  - Preimplantation
OT  - Sexing
COIS- Declaration of Competing Interest ME, MB, HG, BE, PBB and MF declare no conflicts 
      of interest related to this study.
EDAT- 2020/06/27 06:00
MHDA- 2021/05/11 06:00
CRDT- 2020/06/27 06:00
PHST- 2020/01/18 00:00 [received]
PHST- 2020/06/08 00:00 [revised]
PHST- 2020/06/11 00:00 [accepted]
PHST- 2020/06/27 06:00 [pubmed]
PHST- 2021/05/11 06:00 [medline]
PHST- 2020/06/27 06:00 [entrez]
AID - S0301-2115(20)30387-0 [pii]
AID - 10.1016/j.ejogrb.2020.06.023 [doi]
PST - ppublish
SO  - Eur J Obstet Gynecol Reprod Biol. 2020 Sep;252:89-93. doi: 
      10.1016/j.ejogrb.2020.06.023. Epub 2020 Jun 12.