[Preliminary study on prenatal diagnosis from coelomic cells using polymerase chain reaction].
OBJECTIVE
To evaluate the number of cells in coelomic fluid by DNA quantitation with picogreen and to establish a new method of prenatal diagnosis for sex-linked disorders in early pregnancy.


METHODS
Coelomic fluid and chorionic villi were obtained from 25 women undergoing planned termination of pregnancy. DNA of coelomic fluid was extracted by boiling method and DNA concentration was measured with picogreen, and fetal gender was determined by X and Y-sequence-specific PCR. The results of PCR were compared with traditional cytogenetic analysis of chorionic villi.


RESULTS
There were 750-6270 cells in 0.5-3 ml coelomic fluid. Two amplified bands of X and Y were identified simultaneously in the coelomic fluid of male-fetus. Only one band was detected in that of female-fetus. Eleven out of 25 coelomic fluids showed X and Y bands, and the others were one band (X). The overall concordance rate of gender determination between coelomic fluid and chorionic villi was 100%.


CONCLUSIONS
The number of cells in 0.5-3 ml coelomic fluid can completely meet the needs of PCR reactions. Sex determination using cells in coelomic fluid and PCR is reliable, which can be used as a prenatal diagnostic test for sex-linked disorders and has the advantages of high sensitivity and rapidity.
Sun
Lu-ming
LM
Department of Obstetrics, Shanghai First Maternal and Infant Health Hospital, Shanghai 200040, China.
Wang
De-fen
DF
Liu
Jian-hua
JH
D001826:Body Fluids / Q000737:chemistry; D004247:DNA / Q000032:analysis*; D017931:DNA Primers; D005260:Female; D005333:Fetus / Q000166:cytology; D030342:Genetic Diseases, Inborn / Q000175:diagnosis*; D008040:Genetic Linkage; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011296:Prenatal Diagnosis / Q000379:methods*; D012743:Sex Preselection
D017931:DNA Primers; D004247:DNA