| The MHE Research Foundation would like to thank Transgenomic, Inc. for the description of this testing. |
Please note that DHPLC for clinical genetic testing MHE / MO / HME is offered at : Department of
Medical Genetics, University of Antwerp (Belgium) & The Genetic Unit ,The Rizzoli Orthopaedic
Institute, Bologna, Italy and GENDIA
Medical Genetics, University of Antwerp (Belgium) & The Genetic Unit ,The Rizzoli Orthopaedic
Institute, Bologna, Italy and GENDIA
Novel EXT1 and EXT2 mutations identified by DHPLC in Italian patients with multiple
osteochondromas. Full Text PDF Link
Pedrini E, De Luca A, Valente EM, Maini V, Capponcelli S, Mordenti M, Mingarelli R, Sangiorgi L,
Dallapiccola B. Modulo di Familiarita e Genetica, Lab. Ricerca Oncologica, Istituti Ortopedici Rizzoli,
Bologna, Italy.
We describe the results of an optimised DHPLC-based mutation screening of the EXT1 and EXT2
genes in Italian patients affected by multiple osteochondromas [MO; also referred to as hereditary
multiple exostoses (HME) in the literature], using a multistep approach. We first analysed 36
unrelated probands for EXT1 mutations by DHPLC analysis and subsequent direct sequencing of all
samples with abnormal elution profile. Negative cases were then screened for EXT2 mutations using
the same approach. In patients who tested normal at DHPLC screening, all EXT1 and EXT2 exons
and splice-site junctions were directly sequenced. In 7 informative families, we also performed a
pre-screening linkage analysis to selectively focus the DHPLC testing on the EXT1 or EXT2 gene. We
detected 31 MO-related mutations, of which 23 (74%) were novel. Seven polymorphisms were also
found. Twenty-four mutations (77%) were found in EXT1 and 7 (23%) in EXT2. No disease-causing
mutations were detected in five of 36 patients, with a mutation frequency of 86%. According with
previous studies, most mutations (90%) are loss of function. Neither false positive nor false negative
results were obtained. This multistep method can be considered a fast and reliable diagnostic
strategy for the detection of EXT1/2 mutations, with excellent sensitivity and specificity.
osteochondromas. Full Text PDF Link
Pedrini E, De Luca A, Valente EM, Maini V, Capponcelli S, Mordenti M, Mingarelli R, Sangiorgi L,
Dallapiccola B. Modulo di Familiarita e Genetica, Lab. Ricerca Oncologica, Istituti Ortopedici Rizzoli,
Bologna, Italy.
We describe the results of an optimised DHPLC-based mutation screening of the EXT1 and EXT2
genes in Italian patients affected by multiple osteochondromas [MO; also referred to as hereditary
multiple exostoses (HME) in the literature], using a multistep approach. We first analysed 36
unrelated probands for EXT1 mutations by DHPLC analysis and subsequent direct sequencing of all
samples with abnormal elution profile. Negative cases were then screened for EXT2 mutations using
the same approach. In patients who tested normal at DHPLC screening, all EXT1 and EXT2 exons
and splice-site junctions were directly sequenced. In 7 informative families, we also performed a
pre-screening linkage analysis to selectively focus the DHPLC testing on the EXT1 or EXT2 gene. We
detected 31 MO-related mutations, of which 23 (74%) were novel. Seven polymorphisms were also
found. Twenty-four mutations (77%) were found in EXT1 and 7 (23%) in EXT2. No disease-causing
mutations were detected in five of 36 patients, with a mutation frequency of 86%. According with
previous studies, most mutations (90%) are loss of function. Neither false positive nor false negative
results were obtained. This multistep method can be considered a fast and reliable diagnostic
strategy for the detection of EXT1/2 mutations, with excellent sensitivity and specificity.
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| DHPLC Genetic Testing |
(DHPLC) denaturing high performance liquid chromatography: DHPLC is a more recent and a
more accurate and sensitive form of testing than other methods for mutation detection.
Click the link DHPLC below to view an animation of this testing
Press Release 1 / 19 / 08
Mutation Screening of EXT1 and EXT2 by Denaturing High-Performance Liquid Chromatography,
Direct Sequencing Analysis, Fluorescence in Situ Hybridization, and a New Multiplex
Ligation-Dependent Probe Amplification Probe Set in Patients with Multiple Osteochondromas
Ivy Jennes*, Mark M. Entius{dagger}, Els Van Hul*, Alessandro Parra{ddagger}, Luca
Sangiorgi{ddagger} and Wim Wuyts*
To Read this publication Click Here
Both Luca Sangiorgi, M.D., Ph.D. and Wim Wuyts, Ph.D. are members of our Scientific & Medical
Advisory Board and our foundation helped support this research
Mutation Screening of EXT1 and EXT2 by Denaturing High-Performance Liquid Chromatography,
Direct Sequencing Analysis, Fluorescence in Situ Hybridization, and a New Multiplex
Ligation-Dependent Probe Amplification Probe Set in Patients with Multiple Osteochondromas
Ivy Jennes*, Mark M. Entius{dagger}, Els Van Hul*, Alessandro Parra{ddagger}, Luca
Sangiorgi{ddagger} and Wim Wuyts*
To Read this publication Click Here
Both Luca Sangiorgi, M.D., Ph.D. and Wim Wuyts, Ph.D. are members of our Scientific & Medical
Advisory Board and our foundation helped support this research
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By the Health On the Net Foundation. Click here to verify.# HON Conduct 282463 and is linked on the NIH
National Library of Medicine, Directory of Health Organizations (SIS) website, as well as the link for Patient
Information on The Diseases Database a cross-referenced index of human disease, and the Intute: health & life
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located in the UK
By the Health On the Net Foundation. Click here to verify.# HON Conduct 282463 and is linked on the NIH
National Library of Medicine, Directory of Health Organizations (SIS) website, as well as the link for Patient
Information on The Diseases Database a cross-referenced index of human disease, and the Intute: health & life
sciences a free online service providing access to the very best Web resources for education and research
located in the UK
The MHE Research Foundation is proud to be working with the EuroBoNeT consortium, a European Commission
granted Network of Excellence for studying the pathology and genetics of bone tumors.
granted Network of Excellence for studying the pathology and genetics of bone tumors.
