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Clinical Chemistry 52: 2013-2020, 2006. First published September 28, 2006; 10.1373/clinchem.2006.074971
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(Clinical Chemistry. 2006;52:2013-2020.)
© 2006 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

Use of Routinely Collected Amniotic Fluid for Whole-Genome Expression Analysis of Polygenic Disorders

Gyula Richárd Nagy1,a, Balázs Gyõrffy2, Orsolya Galamb3, Béla Molnár3, Bálint Nagy1 and Zoltán Papp1

1 1st Department of Obstetrics and Gynecology, Semmelweis University, Budapest, Hungary.
2 Szentágothai János Knowledge Center, Semmelweis University, Budapest, Hungary.
3 2nd Department of Internal Medicine, Cell Analysis Laboratory, Semmelweis University, Clinical Research Unit of the Hungarian Academy of Sciences, Budapest, Hungary.

aAddress correspondence to this author at: Semmelweis University, Faculty of Medicine, 1st Department of Obstetrics and Gynecology, 27 Baross str., Budapest H-1088, Hungary. Fax 36-1-317-6174; e-mail nagygyr{at}noi1.sote.hu.

Background: Neural tube defects related to polygenic disorders are the second most common birth defects in the world, but no molecular biologic tests are available to analyze the genes involved in the pathomechanism of these disorders. We explored the use of routinely collected amniotic fluid to characterize the differential gene expression profiles of polygenic disorders.

Methods: We used oligonucleotide microarrays to analyze amniotic fluid samples obtained from pregnant women carrying fetuses with neural tube defects diagnosed during ultrasound examination. The control samples were obtained from pregnant women who underwent routine genetic amniocentesis because of advanced maternal age (>35 years). We also investigated specific folate-related genes because maternal periconceptional folic acid supplementation has been found to have a protective effect with respect to neural tube defects.

Results: Fetal mRNA from amniocytes was successfully isolated, amplified, labeled, and hybridized to whole-genome transcript arrays. We detected differential gene expression profiles between cases and controls. Highlighted genes such as SLA, LST1, and BENE might be important in the development of neural tube defects. None of the specific folate-related genes were in the top 100 associated transcripts.

Conclusions: This pilot study demonstrated that a routinely collected amount of amniotic fluid (as small as 6 mL) can provide sufficient RNA to successfully hybridize to expression arrays. Analysis of the differences in fetal gene expressions might help us decipher the complex genetic background of polygenic disorders.






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