Sex determination of human preimplantation embryo using nested polymerase chain reaction.
- Author:
Zhe-xu JIAO
1
;
Guang-lun ZHUANG
;
Can-quan ZHOU
;
Min-fang ZHANG
;
Li-lin LI
Author Information
- Publication Type:Journal Article
- MeSH: Amelogenin; Blastocyst; cytology; metabolism; Blastomeres; cytology; metabolism; DNA; genetics; Dental Enamel Proteins; genetics; Female; Genotype; Humans; Lymphocytes; cytology; metabolism; Male; Polymerase Chain Reaction; methods; Sex Determination Analysis; methods
- From: Chinese Journal of Medical Genetics 2003;20(1):64-65
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEUsing nested polymerase chain reaction (PCR) to perform preimplantation gender diagnosis.
METHODSOne (or two) lymphocyte and blastomere (n=50/group) were collected and prepared under the following conditions: (1) water only (H(2)O); (2) freeze-thaw liquid nitrogen, then boiling; (3) potassium hydroxide/dithiotheriol, heated to 65 degree centigrade, followed by acid neutralization (KOH). Cells were analyzed by PCR using nested primers amplification with amelogenin gene.
RESULTSThe amplification rate and allele dropout (ADO) rate for male lymphocytes by the three methods were 83%, 94%, 95% and 24%, 12%, 4%, respectively. Using two cells per reaction did not increase the amplification rate for the KOH method.
CONCLUSIONThe KOH method for DNA preparation is superior to the other methods evaluated. Dual blastomere biopsy and independent blastomere analysis may improve preimplantation diagnostic reliability.
