Validation of sensitivity and specificity of tetraplet-primed PCR (TP-PCR) in the molecular diagnosis of myotonic dystrophy type 2 (DM2)
| Autor: | Claudio Catalli, Fabrizio Rinaldi, Annalisa Botta, Giuseppe Novelli, Raniero Iraci, Alessandra Morgante |
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| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
musculoskeletal diseases
myotonic disorders polymerase chain reaction Myotonic Disorder Biology medicine.disease_cause Myotonic dystrophy Pathology and Forensic Medicine law.invention agar gel Degenerative disease law Genotype medicine Myotonic Dystrophy Allele humans Polymerase chain reaction Southern blot Electrophoresis Agar Gel Mutation dna primers sensitivity and specificity electrophoresis agar gel molecular diagnostic techniques medicine.disease Molecular biology Settore MED/03 - Genetica Medica electrophoresis Molecular Medicine Regular Articles |
| Popis: | Myotonic dystrophy type 2 (DM2, OMIM #602688) is a multisystemic hereditary degenerative disease caused by a tetranucleotide CCTG expansion in the ZNF9 gene. Routine testing strategies for DM2 require the use of Southern blot or long-range PCR, but the presence of very large expansions and wide somatic mosaicism greatly reduce the sensitivity of these reference techniques. We therefore developed and validated a tetraplet-primed PCR (TP-PCR) method to detect the DM2 mutation by testing 87 DM2-positive and 76 DM2-negative previously characterized patients. The specificity of this technique was evaluated including DNA samples from 39 DM1-positive patients. We then attempted a prospective analysis of 50 patients with unknown genotype who referred to our center for diagnostic or presymptomatic tests. Results show that TP-PCR is a fast, reliable, and flexible technique, whose specificity and sensitivity is almost 100%, with no false positive or negative results either in retrospective and prospective applications. We therefore conclude that using this technique, in combination with the short-range PCR, is sufficient to correctly establish the presence or the absence of ZNF9 expanded alleles in the molecular diagnosis of DM2. |
| Databáze: | OpenAIRE |
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