Accurate, fast and cost-effective diagnostic test for monosomy 1p36 using real-time quantitative PCR.
Monosomy 1p36 is considered the most common subtelomeric deletion syndrome in humans and it accounts for 0.5-0.7% of all the cases of idiopathic intellectual disability. The molecular diagnosis is often made by microarray-based comparative genomic hybridization (aCGH), which has the drawback of being a high-cost technique. However, patients with classic monosomy 1p36 share some typical clinical characteristics that, together with its common prevalence, justify the development of a less expensive, targeted diagnostic method. In this study, we developed a simple, rapid, and inexpensive real-time quantitative PCR (qPCR) assay for targeted diagnosis of monosomy 1p36, easily accessible for low-budget laboratories in developing countries. For this, we have chosen two target genes which are deleted in the majority of patients with monosomy 1p36: PRKCZ and SKI. In total, 39 patients previously diagnosed with monosomy 1p36 by aCGH, fluorescent in situ hybridization (FISH), and/or multiplex ligation-dependent probe amplification (MLPA) all tested positive on our qPCR assay. By simultaneously using these two genes we have been able to detect 1p36 deletions with 100% sensitivity and 100% specificity. We conclude that qPCR of PRKCZ and SKI is a fast and accurate diagnostic test for monosomy 1p36, costing less than 10 US dollars in reagent costs.
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, MG, Brazil.
Pricila da Silva
Pda S
Cunha
GENE-Núcleo de Genética Médica, Avenida Afonso Pena 3111, 9th Floor, Belo Horizonte 30130-909, MG, Brazil.
Heloisa B
HB
Pena
Departamento de Genética e Biologia Evolutiva, Instituto de Biociências, Universidade de São Paulo, São Paulo 05508-900, SP, Brazil.
Carla Sustek
CS
D'Angelo
Departamento de Genética e Biologia Evolutiva, Instituto de Biociências, Universidade de São Paulo, São Paulo 05508-900, SP, Brazil.
Celia P
CP
Koiffmann
Signature Genomic Laboratories, PerkinElmer, Inc., Spokane, WA 99207, USA.
Jill A
JA
Rosenfeld
Paw Print Genetics, Genetic Veterinary Sciences, Inc., Spokane, WA 99202, USA.
Lisa G
LG
Shaffer
GENE-Núcleo de Genética Médica, Avenida Afonso Pena 3111, 9th Floor, Belo Horizonte 30130-909, MG, Brazil.
Martin
M
Stofanko
GENE-Núcleo de Genética Médica, Avenida Afonso Pena 3111, 9th Floor, Belo Horizonte 30130-909, MG, Brazil.
Higgor
H
Gonçalves-Dornelas
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, MG, Brazil ; GENE-Núcleo de Genética Médica, Avenida Afonso Pena 3111, 9th Floor, Belo Horizonte 30130-909, MG, Brazil.
Sérgio Danilo Junho
SD
Pena
D002872:Chromosome Deletion; D002878:Chromosomes, Human, Pair 1 / Q000235:genetics*; D003362:Cost-Benefit Analysis; D005260:Female; D005819:Genetic Markers; D006801:Humans; D008297:Male; D025202:Molecular Diagnostic Techniques / Q000191:economics; D009006:Monosomy / Q000175:diagnosis; D060888:Real-Time Polymerase Chain Reaction / Q000191:economics*
D005819:Genetic Markers