Enhanced Detection and Sizing of the HTT CAG Repeat Expansion in Huntington Disease Using an Improved Triplet-Primed PCR Assay.
Neurodegener Dis
; 16(5-6): 348-51, 2016.
Article
en En
| MEDLINE
| ID: mdl-27207688
ABSTRACT
BACKGROUND:
Accurate determination of the CAG repeat number is crucial for diagnostic and predictive testing for Huntington disease (HD). Current PCR-based assays can accurately size up to â¼110 HTT CAG repeats.OBJECTIVE:
To develop an improved assay capable of detecting larger CAG repeat expansions.METHODS:
A triplet-primed PCR (TP-PCR) assay was optimized and validated on 14 HD reference DNAs, including a sample carrying a large expansion of â¼180 CAG repeats.RESULTS:
All 14 HD reference samples showed 100% concordance with the previously verified allele sizes. For alleles under 45 CAGs, identical repeat sizes were obtained, while alleles larger than 46 CAGs were sized to within ±1 CAG. The improved TP-PCR assay successfully detected the â¼180 CAG repeat allele in an affected sample.CONCLUSION:
This method extends the detection limit of large expanded alleles to at least â¼175-180 CAG repeats, thus reducing the likelihood of requiring Southern blot analysis for any HD-affected sample.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Reacción en Cadena de la Polimerasa
/
Enfermedad de Huntington
/
Expansión de Repetición de Trinucleótido
/
Proteína Huntingtina
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
Límite:
Humans
Idioma:
En
Revista:
Neurodegener Dis
Asunto de la revista:
NEUROLOGIA
Año:
2016
Tipo del documento:
Article
País de afiliación:
Singapur