Clinical Chemistry
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Clinical Chemistry 47: 645-653, 2001;
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(Clinical Chemistry. 2001;47:645-653.)
© 2001 American Association for Clinical Chemistry, Inc.

Articles

{zeta}-, {epsilon}-, and {gamma}-Globin mRNA in Blood Samples and CD71+ Cell Fractions from Fetuses and from Pregnant and Nonpregnant Women, with Special Attention to Identification of Fetal Erythroblasts

Anne Mette Høgh1,1, Thomas Vauvert F. Hviid1,1,a, Britta Christensen3, Steen Sørensen1, Rasmus D. Larsen5, Steen Smidt-Jensen2, Jens Bang4 and John Philip3

1 Departments of Clinical Biochemistry 339, and
2 Gynecology and Obstetrics, Copenhagen University Hospital, H:S Hvidovre Hospital, 30 Kettegaard Allé, DK-2650 Hvidovre, Denmark.

3 The Chromosome Laboratory, Prenatal Research Unit, and
4 Department of Obstetrics and Gynecology, Juliane Marie Center, H:S Rigshospitalet, University of Copenhagen, 9 Blegdamsvej, DK-2100 Copenhagen Ø, Denmark.

5 Dako A/S, 42 Produktionsvej, DK-2600 Glostrup, Denmark.
a Author for correspondence. Fax 45-3675-0977; email thomas.hviid{at}hh.hosp.dk or hviid{at}dadlnet.dk.

Background: Information about the appearance of {gamma}-, {epsilon}-, and {zeta}-globin mRNAs in fetal erythroblasts during gestation and about the presence and amounts of these mRNAs in pregnant and nonpregnant women is important from the perspective of using these molecules as a marker of fetal erythroblasts. A specific marker is necessary for isolation and identification of fetal nucleated red blood cells from maternal blood samples for use in antenatal diagnosis of fetal genetic or chromosomal abnormalities.

Methods: We used a very sensitive reverse transcription-PCR (RT-PCR) method, coamplification analysis of {gamma}- and {epsilon}-globin cDNA, and quantitative analysis of {gamma}-globin mRNA based on competitive RT-PCR to investigate these aspects.

Results: All adult whole-blood samples were negative for {epsilon}- and {zeta}-globin mRNA. Analyses of CD71+ cell fractions showed that specimens from 19 of 20 nonpregnant and 10 of 14 pregnant women (at 9–13 weeks of gestation) were positive for {gamma}-globin mRNA (Fisher’s exact test, P = 0.13), and those from 3 of 20 nonpregnant and 5 of 14 pregnant women were positive for {zeta}-globin mRNA (Fisher’s exact test, P = 0.23). No {epsilon}-globin mRNA was detected in CD71+ cell fractions from 1-mL blood samples from adults. CD71+ cell fractions from eight fetal blood samples (at 17–20 weeks of gestation) were positive for all three globin mRNAs. We found no statistically significant difference between the amounts of {gamma}-globin mRNA in pregnant and nonpregnant women.

Conclusions: This study indicates that {epsilon}-globin mRNA might function as a marker for fetal CD71+ cells early in pregnancy. Although {gamma}-globin mRNA can be detected in CD71+ cell fractions from most adults, these transcripts also may be of use because of a marked difference between adult and fetal values.




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 Mol Hum ReprodHome page
M. Choolani, K. O'Donoghue, D. Talbert, S. Kumar, I. Roberts, E. Letsky, P. R. Bennett, and N. M. Fisk
Characterization of first trimester fetal erythroblasts for non-invasive prenatal diagnosis
Mol. Hum. Reprod., April 1, 2003; 9(4): 227 - 235.
[Abstract] [Full Text] [PDF]

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 Clin. Chem.Home page
T. V. Hviid
In-Cell PCR Method for Specific Genotyping of Genomic DNA from One Individual in a Mixture of Cells from Two Individuals: A Model Study with Specific Relevance to Prenatal Diagnosis Based on Fetal Cells in Maternal Blood
Clin. Chem., December 1, 2002; 48(12): 2115 - 2123.
[Abstract] [Full Text] [PDF]


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