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Detection of Hereditary Persistence of -Fetoprotein by Conformation-sensitive Gel Electrophoresis Analysis

¹ Fundación Valenciana, de Investigaciones Biomédicas, Instituto de Investigaciones Citológicas, Amadeo de Saboya, 4, 46010 Valencia, Spain
² Hospital "Arnau de Vilanova", San Clemente 12, 46015 Valencia, Spain

^aAddress correspondence to this author at: Fundación Valenciana de Investigaciones Biomédicas, Laboratory of Cell Organization, Amadeo de Saboya, 4, 46010 Valencia, Spain. Fax 34-963601453; e-mail hernan{at}ochoa.fib.es.

To the Editor:

The concentration of human -fetoprotein (AFP; MIM 104150) is used as a marker for several diseases in both adults and children, and it is also systematically measured in pregnant women as a marker of defects in the fetus.

In 1983, a case was discovered of a woman with high AFP expression that was shown to be attributable to a benign autosomal dominant genetic trait, hereditary persistence of AFP (HPAFP) (1). This syndrome shows high expression of AFP, but there is no association with any pathology. Failure to recognize HPAFP can lead to unsuitable treatments. For example, a 20-month-old child with HPAFP underwent surgery for a testicular germ cell tumor (2).

In one family, all affected members exhibited an identical haplotype: a heterozygous GA substitution (3) at position -116 of the 5'-flanking region of the AFP gene. The same haplotype was also reported in another HPAFP family (4). The frequency of this anomaly is unknown because of the paucity of reported cases. As it is an autosomal dominant genetic trait, the routine inclusion of a test for HPAFP should be seriously considered in pregnant women and patients with high AFP concentrations to prevent inappropriate treatment decisions.

We show that conformation-sensitive gel electrophoresis (CSGE) can detect the GA mutation in a possibly affected case of HPAFP.

Seventeen DNA samples from members of a family with HPAFP (4), with their mutational analyses carried out by sequencing in our laboratory, were used in this study. This familial study was performed with the formal consent of all members of the family, who were previously informed of the aim of the study.

The promoter region of interest was amplified from genomic DNA; the PCR product was 260 bp in length. The PCR amplifications were done as indicated in the Data Supplement that accompanies the online version of this letter at http://www.clinchem.org/content/vol49/issue12/. The conditions for CSGE analysis were essentially the same as described previously (5). The electrophoresis was performed in a SQ3 standard manual sequencer (Amersham Biosciences).

The results of the CSGE analysis of six members of the HPAFP family are shown, according to pedigree, in Fig. 1 . Aberrant bands in the DNA samples corresponded to affected members, whereas only one band was obtained from unaffected members. The extra band corresponded to the presence of the GA mutation located in the promoter region of the AFP gene at -116 bp.

View larger version (36K): [in this window] [in a new window]   Figure 1. CSGE results for six members of the HPAFP family, two with normal concentrations of AFP and four with increased AFP. (A), pedigree of the HPAFP family showing the first and second generations. a, age. The numbers below age indicate the AFP concentrations (µg/L). Arrow indicates the proband. The AFP concentration shown for the proband is the mean of five assays. (B), CSGE analysis of AFP PCR products. The unaffected members show one band (a), whereas the affected members show two bands corresponding to the homoduplex (a) and heteroduplex (b) forms. The bands in the samples of the affected members show lower intensities than the single band from unaffected members, indicating the presence of hetero- and homoduplex forms.

To exclude the HPAFP trait in those patients with high AFP, we propose the application of CSGE to rapidly and efficiently detect the heterozygous GA substitution in the AFP gene.

Acknowledgments

This research was supported in part by the Fondo de Investigación Sanitaria (FIS PI021345). We are most grateful to the proband and her relatives for generously participating in this research. Dr. J. R. Blesa is an associate researcher of the Fundación Valenciana de Investigaciones Biomédicas (FVIB). M. L. Lacalle is a predoctoral fellow of FVIB.

References

1. Ferguson-Smith MA, May HM, O’Hare E, Aitken DA. Hereditary persistence of -fetoprotein: a new autosomal dominant trait identified in an antenatal screening programme for spina bifida. J Med Genet 1983;20:458.
2. Cochran PK, Chauvenet AR, Hart PS, de Graaf SS, Cushing B, Kroovand L, et al. Hereditary persistence of -fetoprotein in a child with testicular germ cell tumor. Med Pediatr Oncol 1999;32:436-437.[CrossRef][ISI][Medline] [Order article via Infotrieve]
3. McVey JH, Michaelides K, Hansen LP, Ferguson-Smith M, Tilghman S, Krumlauf R, et al. A GA substitution in an HNF I binding site in the human -fetoprotein gene is associated with hereditary persistence of -fetoprotein (HPAFP). Hum Mol Genet 1993;2:379-384.[Abstract/Free Full Text]
4. Blesa JR, Giner-Durán R, Vidal J, Lacalle ML, Catalan I, Bixquert M, et al. Report of hereditary persistence of -fetoprotein in a Spanish family: molecular basis and clinical concerns. J Hepatol 2003;38:541-544.[CrossRef][ISI][Medline] [Order article via Infotrieve]
5. Körkko J, Annunen S, Pihlajama T, Prockop DJ, Ala-kokko L. Conformation sensitive gel electrophoresis for simple and accurate detection of mutations: comparison with denaturing gradient gel electrophoresis and nucleotide sequencing. Proc Natl Acad Sci U S A 1998;95:1681-1685.[Abstract/Free Full Text]

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This Article Extract Full Text (PDF) Supplemental Data Submit an electronic Letter to the Editor about this paper Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (1) Citing Articles via Google Scholar Google Scholar Articles by Blesa, J. R. Articles by Hernández-Yago, J. Search for Related Content PubMed PubMed Citation Articles by Blesa, J. R. Articles by Hernández-Yago, J. Related Collections Molecular Diagnostics and Genetics