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201904151249411

PHP1b-QAMAPCR real time    Product information

Quantification assay of PHP1b                  cod. BM-001

 

Principle of the test : Quantification of altered methylation after bisulfite conversion of DNA

Technology : QAMAPCR real time

Gene Target : GNAS

Specimen : DNA

Results within : Pathological range IM < 30

Reporting Units : IM Index of Methylation

Number of tests : 25 tests BM-001 

Kit storage : -20°C

Necessary equipment : MethylEdge Bisulfite Conversion System, Thermocycler, 7500 Real Time PCR System

Status : Ready to use

PHP1b-QAMAPCR real time cod. BM-001

 

Quantification assay of pseudohypoparathyroidism type 1b (PHP1b)

 

·      PHP1b-QAMA real time Quantification complete kit 25 tests BM-001

 

Altered methylation patterns have been found to play a role in developmental disorders, cancer and aging. Increasingly, changes in DNA methylation are used as molecular markers of disease. Pathogenesis of pseudohypoparathyroidism type 1b (PHP-1b) is related to the loss of methylation at the GNAS exon A/B region, which is combined with epigenetic defects at other differentially methylated GNAS regions in most sporadic cases. Originally PHP-1a and 1b were classified only by clinical symptoms. Molecular diagnosis helped us to distinguish two types of PHP, but genetic and epigenetic defect doesn’t always correspond to specific synptoms. The distinction between PHP-1a and 1b remains unclear. We believe the most important information for PHP patients is about inheritance, and our method must contribute to patients to give rapid information about it 1,2.

 

We have established a novel quantitative analysis of methylated alleles (QAMA) which is essentially a major improvement over a previous method based on real-time PCR (MethyLight)3. This method is based on real-time PCR on bisulfite-treated DNA. From the genomic sequence of GNAS (GenBank accession number AL121917) exon A/B region, DNA sequences of MET and UNMET after bisulfite modification were deduced. The PHP1b-QAMAPCR real time kit is designed to measure DNA methylation index (IM) of exon A/B region GNAS gene by QAMA real-time PCR after DNA extraction and bisulfite conversion. User friendly and complete, the PHP1b-QAMAPCR real time kit is suitable for any laboratory.

 

Reference

1.      Kinoshita K, Minagawa M, Takatani T, Takatani R, Ohashi M, Kohno Y. Establishment of diagnosis by bisulfite-treated methylation-specific PCR method and analysis of clinical characteristics of pseudohypoparathyroidism type 1b. Endocr J. 2011;58(10):879-87.

2.      Elli FM, de Sanctis L, Bollati V, Tarantini L, Filopanti M, Barbieri AM, Peverelli E, Beck-Peccoz P, Spada A, Mantovani G. Quantitative analysis of methylation defects and correlation with clinical characteristics in patients with pseudohypoparathyroidism type I and GNAS epigenetic alterations. J Clin Endocrinol Metab. 2014 Mar;99(3):E508-17. doi: 10.1210/jc.2013-3086.

3.      Zeschnigk M, Böhringer S, Price EA, Onadim Z, Masshöfer L, Lohmann DR. A novel real-time PCR assay for quantitative analysis of methylated alleles (QAMA): analysis of the retinoblastoma locus. Nucleic Acids Res. 2004 Sep 7;32(16):e125.