Simple, rapid and inexpensive quantitative fluorescent PCR method for detection of microdeletion and microduplication syndromes

PLoS One. 2013 Apr 19;8(4):e61328. doi: 10.1371/journal.pone.0061328. Print 2013.

Abstract

Because of economic limitations, the cost-effective diagnosis of patients affected with rare microdeletion or microduplication syndromes is a challenge in developing countries. Here we report a sensitive, rapid, and affordable detection method that we have called Microdeletion/Microduplication Quantitative Fluorescent PCR (MQF-PCR). Our procedure is based on the finding of genomic regions with high homology to segments of the critical microdeletion/microduplication region. PCR amplification of both using the same primer pair, establishes competitive kinetics and relative quantification of amplicons, as happens in microsatellite-based Quantitative Fluorescence PCR. We used patients with two common microdeletion syndromes, the Williams-Beuren syndrome (7q11.23 microdeletion) and the 22q11.2 microdeletion syndromes and discovered that MQF-PCR could detect both with 100% sensitivity and 100% specificity. Additionally, we demonstrated that the same principle could be reliably used for detection of microduplication syndromes, by using patients with the Lubs (MECP2 duplication) syndrome and the 17q11.2 microduplication involving the NF1 gene. We propose that MQF-PCR is a useful procedure for laboratory confirmation of the clinical diagnosis of microdeletion/microduplication syndromes, ideally suited for use in developing countries, but having general applicability as well.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Androgen-Insensitivity Syndrome / diagnosis*
  • Androgen-Insensitivity Syndrome / genetics
  • Chromosome Deletion*
  • Chromosome Duplication / genetics*
  • DiGeorge Syndrome / diagnosis*
  • DiGeorge Syndrome / genetics
  • Fluorescence
  • Humans
  • Methyl-CpG-Binding Protein 2 / genetics
  • Polymerase Chain Reaction / economics*
  • Polymerase Chain Reaction / methods*
  • Williams Syndrome / diagnosis*
  • Williams Syndrome / genetics

Substances

  • MECP2 protein, human
  • Methyl-CpG-Binding Protein 2

Supplementary concepts

  • Lubs syndrome

Grants and funding

The authors thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico of Brazil (CNPq) for its support of this Project 401983/2010-2. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.