title: Validation of Polymerase Chain Reaction-Based Assay to Detect Actual Number of CGG Repeats in FMR1 Gene in Indian Fragile X Syndrome Patients.
abstract: Molecular genetic testing for fragile X (FX) is complicated due to the large variation in the size of CGG expansion. The aim of this study was to apply this new technique using AmplideX FMR1 PCR assay, which is considered a better diagnostic tool for detecting expanded alleles in Indian population. The primary objective was to identify the carrier status of females and to correlate the instability of premutation alleles in females with the repeat sizes. 24 children with FX based on rapid PCR and 29 female relatives of these patients were included. Out of the 29 females screened, those whose child (or children) was affected by FX, were all premutation carriers confirming their role in transmission. The smallest PM allele that expanded into FM in the next generation was 78 repeats and the smallest PM allele detected was 63 repeats, and when transmitted from mother to offspring remained in the premutation range. In 4 families, the repeat size of the allele reduced from PM to normal repeat numbers in their daughters and in 1 case to borderline PM range. Thus, apart from the reduced turnaround time, this PCR based assay offers advantage by its sensitivity to detect CGG repeats in the intermediate region and lower range of premutation alleles. It also provides added information of AGG interruptions, which may have an impact on the counseling of women with intermediate and PM alleles.
authors.affiliation: 1 Genetics Division, Department of Pediatrics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Madhumita Roy
authors.initials: MR
authors.lastname: Chowdhury
authors.affiliation: 1 Genetics Division, Department of Pediatrics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Sandeepa
authors.initials: S
authors.lastname: Chauhan
authors.affiliation: 2 Department of Genetics, University of Delhi, South Campus, New Delhi, India.
authors.forename: Anjali
authors.initials: A
authors.lastname: Dabral
authors.affiliation: 2 Department of Genetics, University of Delhi, South Campus, New Delhi, India.
authors.forename: B K
authors.initials: BK
authors.lastname: Thelma
authors.affiliation: 1 Genetics Division, Department of Pediatrics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Neerja
authors.initials: N
authors.lastname: Gupta
authors.affiliation: 1 Genetics Division, Department of Pediatrics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Madhulika
authors.initials: M
authors.lastname: Kabra
mesh_terms: D044466:Asian People / Q000235:genetics; D015139:Blotting, Southern; D005190:Family; D005260:Female; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000235:genetics*; D005600:Fragile X Syndrome / Q000235:genetics*; D005817:Genetic Counseling; D020022:Genetic Predisposition to Disease; D006801:Humans; D007194:India; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D019680:Trinucleotide Repeat Expansion
chemical_list: D051860:Fragile X Messenger Ribonucleoprotein 1; C497683:FMR1 protein, human
keywords: AGG interruption; CGG repeats; full mutation; intermediate zone; premutation