Fragile X protein in newborn dried blood spots

BMC Medical Genetics

W Ted Brown¹  Nicole Hosmer¹  Anne Glicksman¹  Sarah L Nolin¹  Richard Kascsak⁵  Tiffany Wotton²  Michael Field³  Veronica Wiley²  Michele Caggana⁴  Carl Dobkin¹  Giuseppe LaFauci⁵  Tatyana Adayev⁵ 

[1] Department of Human Genetics, New York State Institute for Basic Research in Developmental Disabilities, 1050 Forest Hill Road, Staten Island, New York 10314, USA;NSW Newborn Screening Programme & University of Sydney, Sydney, Australia;The NSW GOLD Service, Hunter Genetics, Newcastle, Australia;Wadsworth Center, New York State Department of Health, Albany, New York, USA;Department of Developmental Biochemistry, New York State Institute for Basic Research in Developmental Disabilities, 1050 Forest Hill Road, Staten Island, New York 10314, USA

关键词: Newborn screening;    Luminex;    CGG repeats;    Capture immune assay;    DBS;    Dried blood spots;    FMRP;    FMR1;    Fragile X syndrome;   

Others  :  1090299
DOI  :  10.1186/s12881-014-0119-0

 received in 2014-07-31, accepted in 2014-10-13,  发布年份 2014

【 摘 要 】

Background

The fragile X syndrome (FXS) results from mutation of the FMR1 gene that prevents expression of its gene product, FMRP. We previously characterized 215 dried blood spots (DBS) representing different FMR1 genotypes and ages with a Luminex-based immunoassay (qFMRP). We found variable FMRP levels in the normal samples and identified affected males by the drastic reduction of FMRP.

Methods

Here, to establish the variability of expression of FMRP in a larger random population we quantified FMRP in 2,000 anonymous fresh newborn DBS. We also evaluated the effect of long term storage on qFMRP by retrospectively assaying 74 aged newborn DBS that had been stored for 7-84 months that included normal and full mutation individuals. These analyses were performed on 3 mm DBS disks. To identify the alleles associated with the lowest FMRP levels in the fresh DBS, we analyzed the DNA in the samples that were more than two standard deviations below the mean.

Results

Analysis of the fresh newborn DBS revealed a broad distribution of FMRP with a mean approximately 7-fold higher than that we previously reported for fresh DBS in normal adults and no samples whose FMRP level indicated FXS. DNA analysis of the lowest FMRP DBS showed that this was the low extreme of the normal range and included a female carrying a 165 CGG repeat premutation. In the retrospective study of aged newborn DBS, the FMRP mean of the normal samples was less than 30% of the mean of the fresh DBS. Despite the degraded signal from these aged DBS, qFMRP identified the FXS individuals.

Conclusions

The assay showed that newborn DBS contain high levels of FMRP that will allow identification of males and potentially females, affected by FXS. The assay is also an effective screening tool for aged DBS stored for up to four years.

【 授权许可】

   
2014 Adayev et al.; licensee BioMed Central Ltd.

【 图 表 】

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