Prenatal genetic diagnosis from maternal blood: simultaneous immunophenotyping and FISH of fetal nucleated erythrocytes isolated by negative and positive magnetic activated cell sorting.
10.3349/ymj.2000.41.2.258
- Author:
Young Ho YANG
1
;
Kwan Ja JEE
;
Sei Kwang KIM
;
Yong Won PARK
;
In Kyu KIM
;
Dong Hyun CHA
;
Jae Eun CHUNG
;
Sung Hoon KIM
Author Information
1. Department of Obstetrics and Gynecology, Yonsei University College of Medicine, Seoul, Korea. obgy@yumc.yonsei.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Prenatal diagnosis;
fetal nRBC;
density gradient;
MACS;
GPA-immuno FISH
- MeSH:
Erythrocytes/immunology*;
Female;
Fetal Blood/immunology*;
Gestational Age;
Glycophorin;
Human;
Immunomagnetic Separation;
Immunophenotyping;
In Situ Hybridization, Fluorescence*;
Pregnancy;
Prenatal Diagnosis*
- From:Yonsei Medical Journal
2000;41(2):258-265
- CountryRepublic of Korea
- Language:English
-
Abstract:
Fetal nucleated red blood cells (nRBCs) are rare in maternal circulation, but their presence constitutes a potential source of non-invasive prenatal genetic diagnosis. This study was undertaken to establish a non-invasive prenatal genetic diagnosis method using isolated fetal nRBCs. A multi-step method including triple density gradient and magnetic activated cell sorting (MACS) using CD45 and CD71, cytospin centrifugation, K-B staining, and glycophorin A-immuno fluorescence in situ hybridization (GPA-immuno FISH) was performed. The study population included 65 patients from 8 to 41 weeks of gestation, and fetal nRBC was separated from all cases. The number of fetal nRBCs retrieved was 12.8 +/- 2.7 in 8 to 11 gestational weeks, 15.2 +/- 6.5 in 12 to 18 gestational weeks, 16.4 +/- 6.5 in 19 to 23 gestational weeks, 10.6 +/- 3.2 in 24 to 28 gestational weeks, and 5.5 +/- 1.9 in 35 to 41 gestational weeks: the mean number of nRBCs collected from 20 ml of maternal peripheral blood was 13.7 +/- 6.2. The highest value of yield was 45.6% from 12 to 18 weeks gestation. The fetal sex determination confirmed by amniocentesis or chorionic villus sampling showed 100% sensitivity and 91.7% specificity for males; 91.7% sensitivity and 100% specificity for females. We showed that fetal cells can be reliably enriched from maternal blood and that they can be used for detecting specific chromosomes by FISH with a specificity superior to current non-invasive methods.
