PMID- 24078563
OWN - NLM
STAT- MEDLINE
DCOM- 20140102
LR  - 20191210
IS  - 1003-9406 (Print)
IS  - 1003-9406 (Linking)
VI  - 30
IP  - 5
DP  - 2013 Oct
TI  - [Efficiency of multi-round fluorescence in situ hybridization and its influencing 
      factors in preimplantation genetic diagnosis].
PG  - 522-7
LID - 10.3760/cma.j.issn.1003-9406.2013.05.004 [doi]
AB  - OBJECTIVE: To investigate the efficiency of multi-round fluorescence in situ 
      hybridization (FISH) and its influencing factors in preimplantation genetic 
      diagnosis (PGD). METHODS: A total of 48 couples accepted PGD because of various 
      reasons: 24 with Robertsonian translocations, 16 with reciprocal translocations, 
      2 with pericentric inversions, one with advanced maternal age who had a previous 
      liveborn of Down syndrome, 3 suffered from sex chromosome abnormalities and 2 
      repeated spontaneous miscarriages. After 72 retrieval cycles, 432 cleavage stage 
      embryos with more than six cells were biopsied on day three. Only intact nuclei 
      (396) were hybridized in order to verify the chromosomal status of the individual 
      embryos. If previous FISH has failed to give conclusive results while the nuclei 
      remained undamaged, the nuclei were hybridized once again. A total of 870 times 
      of hybridization were conducted to 396 nuclei. Signal identification rates of 
      each round as well as the influence of different probes to the hybridization 
      efficiency were compared. Factors leading to inconclusive FISH results were 
      analyzed as well. RESULTS: Five hundred and thirty five out of 870 hybridizations 
      gave identifiable signals (61.5%). The second and third round FISH showed the 
      best signals with an identification rate of 71.8% and 77.4%, respectively, which 
      were significantly higher than those of the first round (52.8%, P < 0.01), the 
      fourth round (55.8%, P < 0.05, P < 0.01), the fifth round (54.5%, P < 0.05) and 
      the sixth round (27.3%, P < 0.01). The identification rate of centromere specific 
      probe signals (CEP group) was 80.3% and the former three rounds in this group got 
      the best quality of signals with an identification rate of 85.7%, 85.1% and 
      88.0%, respectively, which was significantly higher than that of the latter three 
      rounds. The identification rate of other probe was much lower than with the CEP 
      probe (55.2% vs. 80.3%, P < 0.01) and the best quality of signal in this group 
      was achieved in the fifth round (72.7%), followed by the second round (66.1%) and 
      the third round (63.8%). The identification rate of the first round (50.3%) and 
      the sixth round (22.2%) were significantly lower compared with the second round 
      (P < 0.01). During the 6 rounds of FISH, 335 hybridizations did not give 
      conclusion results (38.5%, 335/870). The main cause of unidentification was weak 
      signals (20.9%, 182/870). Other common factors included background interference 
      (7.6%, 66/870) and failed hybridization (6.1%, 53/870). Rare causes included 
      nucleus damage (1.8%, 16/870), nucleus loss (1.1%, 10/870) and signal 
      split/overlap (0.9%, 8/870). CONCLUSION: Multi-round FISH can improve the utility 
      of single nucleus in PGD and the former three rounds have the highest efficiency. 
      The hybridization effect of CEP is better than other probe. Poor signal quality 
      is the common cause of unidentification results.
FAU - Zhang, Yue-ping
AU  - Zhang YP
AD  - Shanghai JiAi Genetics and IVF Institute, Obstetrics and Gynecology Hospital, 
      Fudan University, Shanghai 200011, P. R. China. Email: steven3019@hotmail.com.
FAU - Zhu, Sai-juan
AU  - Zhu SJ
FAU - Li, Su-ying
AU  - Li SY
FAU - Cao, Xiang
AU  - Cao X
FAU - Sun, Xiao-xi
AU  - Sun XX
LA  - chi
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Zhonghua Yi Xue Yi Chuan Xue Za Zhi
JT  - Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese 
      journal of medical genetics
JID - 9425197
SB  - IM
MH  - Female
MH  - Genetic Testing/*methods
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Male
MH  - Pregnancy
MH  - Preimplantation Diagnosis/*methods
MH  - Prenatal Diagnosis
MH  - Translocation, Genetic
EDAT- 2013/10/01 06:00
MHDA- 2014/01/03 06:00
CRDT- 2013/10/01 06:00
PHST- 2013/10/01 06:00 [entrez]
PHST- 2013/10/01 06:00 [pubmed]
PHST- 2014/01/03 06:00 [medline]
AID - 940630115 [pii]
AID - 10.3760/cma.j.issn.1003-9406.2013.05.004 [doi]
PST - ppublish
SO  - Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2013 Oct;30(5):522-7. doi: 
      10.3760/cma.j.issn.1003-9406.2013.05.004.