<abstract xmlns="http://www.w3.org/1999/xhtml"><bold>Purpose</bold>: Our goal was to develop two-tier strategy based on <italic>in house</italic>-designed methylation specific-duplex polymerase chain reactions (MS-PCRs) that could serve as a relatively simple, cost effective, time efficient approach for molecular screening of imprinted regions on chromosomes 15 and 14.<bold>Patients and methods</bold>: Patients were referred to examination during infancy due to hypotonia and motor development delay. Duplex MS-PCRs were performed that enabled detection of methylated/unmethylated DNA in <italic>NDN</italic> and <italic>MEG3</italic> CpG islands via plurality of detection channels on PCR instrument Rotor Gene 6000.<bold>Results and discussion</bold>: Both, copy number variations as well as methylation changes, were revealed by our <italic>in house-</italic>designed methodology by focusing <italic>NDN</italic> gene. No imprinting aberrations were yet discovered in <italic>MEG3</italic> gene. Clinical features of the patients were compared. In agree with literature no typical facial features were observed in PWS patient with imprinting defect and AS UPD patient seems to have a relatively better development and language ability in comparison to deletional form of the disease.<bold>Conclusion</bold>: In conclusion we were able to establish new, throughput and robust diagnostic approach to PWS/AS.</abstract>

Purpose: Our goal was to develop two-tier strategy based on in house-designed methylation specific-duplex polymerase chain reactions (MS-PCRs) that could serve as a relatively simple, cost effective, time efficient approach for molecular screening of imprinted regions on chromosomes 15 and 14.Patients and methods: Patients were referred to examination during infancy due to hypotonia and motor development delay. Duplex MS-PCRs were performed that enabled detection of methylated/unmethylated DNA in NDN and MEG3 CpG islands via plurality of detection channels on PCR instrument Rotor Gene 6000.Results and discussion: Both, copy number variations as well as methylation changes, were revealed by our in house-designed methodology by focusing NDN gene. No imprinting aberrations were yet discovered in MEG3 gene. Clinical features of the patients were compared. In agree with literature no typical facial features were observed in PWS patient with imprinting defect and AS UPD patient seems to have a relatively better development and language ability in comparison to deletional form of the disease.Conclusion: In conclusion we were able to establish new, throughput and robust diagnostic approach to PWS/AS.

Duplex Methylation-Specific Semi-Quantitative Real-Time PCR for Cost-Effective &amp; Time-Efficient Diagnostic Screening of Chromosome 15 and 14 Imprinted Regions

Department of Clinical Genetics, University Hospital in Hradec Kralove

Institute for Clinical Biochemistry and Diagnostics, University Hospital in Hradec Kralove

Department of Pediatric Neurology, University Hospital in Hradec Kralove, Czech Republic

Acta Medica Martiniana

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Duplex Methylation-Specific Semi-Quantitative Real-Time PCR for Cost-Effective & Time-Efficient Diagnostic Screening of Chromosome 15 and 14 Imprinted Regions

Purpose: Our goal was to develop two-tier strategy based on in house-designed methylation specific-duplex polymerase chain reactions (MS-PCRs) that could serve...

Duplex Methylation-Specific Semi-Quantitative Real-Time PCR for Cost-Effective & Time-Efficient Diagnostic Screening of Chromosome 15 and 14 Imprinted Regions

Data publikacji: 22 sty 2016

Zakres stron: 5 - 12

Otrzymano: 08 wrz 2015

Przyjęty: 20 paź 2015

DOI: https://doi.org/10.1515/acm-2015-0012

Słowa kluczowe
Prader Willi syndrome, Angelman syndrome, methyl-specific semi-quantitative real-time PCR, epigenetics

© 2015 E Ruszova et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Duplex Methylation-Specific Semi-Quantitative Real-Time PCR for Cost-Effective & Time-Efficient Diagnostic Screening of Chromosome 15 and 14 Imprinted Regions

Data publikacji: 22 sty 2016

Zakres stron: 5 - 12

Otrzymano: 08 wrz 2015

Przyjęty: 20 paź 2015

DOI: https://doi.org/10.1515/acm-2015-0012

Słowa kluczowePrader Willi syndrome, Angelman syndrome, methyl-specific semi-quantitative real-time PCR, epigenetics

© 2015 E Ruszova et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Słowa kluczowe
Prader Willi syndrome, Angelman syndrome, methyl-specific semi-quantitative real-time PCR, epigenetics