title: Detection of subtelomere imbalance using MLPA: validation, development of an analysis protocol, and application in a diagnostic centre.
abstract: BACKGROUND
  Commercial MLPA kits (MRC-Holland) are available for detecting imbalance at the subtelomere regions of chromosomes; each kit consists of one probe for each subtelomere.
  
  
  METHODS
  For validation of the kits, 208 patients were tested, of which 128 were known to be abnormal, corresponding to 8528 genomic regions overall. Validation samples included those with trisomy 13, 18 and 21, microscopically visible terminal deletions and duplications, sex chromosome abnormalities and submicroscopic abnormalities identified by multiprobe FISH. A robust and sensitive analysis system was developed to allow accurate interpretation of single probe results, which is essential as breakpoints may occur between MLPA probes.
  
  
  RESULTS
  The validation results showed that MLPA is a highly efficient technique for medium-throughput screening for subtelomere imbalance, with 95% confidence intervals for positive and negative predictive accuracies of 0.951-0.996 and 0.9996-1 respectively. A diagnostic testing strategy was established for subtelomere MLPA and any subsequent follow-up tests that may be required. The efficacy of this approach was demonstrated during 15 months of diagnostic testing when 455 patients were tested and 27 (5.9%) abnormal cases were detected.
  
  
  CONCLUSIONS
  The development of a robust, medium-throughput analysis system for the interpretation of results from subtelomere assays will be of benefit to other Centres wishing to implement such an MLPA-based service.
authors.affiliation: Cytogenetics Department, Guy's and St Thomas' NHS Foundation Trust, London, UK. caroline.ogilvie@genetics.kcl.ac.uk
authors.forename: Joo Wook
authors.initials: JW
authors.lastname: Ahn
authors.forename: Caroline Mackie
authors.initials: CM
authors.lastname: Ogilvie
authors.forename: Alysia
authors.initials: A
authors.lastname: Welch
authors.forename: Helen
authors.initials: H
authors.lastname: Thomas
authors.forename: Rajiv
authors.initials: R
authors.lastname: Madula
authors.forename: Alison
authors.initials: A
authors.lastname: Hills
authors.forename: Celia
authors.initials: C
authors.lastname: Donaghue
authors.forename: Kathy
authors.initials: K
authors.lastname: Mann
mesh_terms: D002648:Child; D002658:Developmental Disabilities / Q000175:diagnosis / Q000235:genetics*; D005820:Genetic Testing; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008607:Intellectual Disability / Q000175:diagnosis / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D016615:Telomere / Q000235:genetics*; D014314:Trisomy / Q000175:diagnosis / Q000235:genetics