Accurate, Fast and Cost-Effective Diagnostic Test for Monosomy 1p36 Using Real-Time Quantitative PCR

被引:3
|
作者
Cunha, Pricila da Silva [1 ]
Pena, Heloisa B. [2 ]
D'Angelo, Carla Sustek [3 ]
Koiffmann, Celia P. [3 ]
Rosenfeld, Jill A. [4 ]
Shaffer, Lisa G. [5 ]
Stofanko, Martin [2 ]
Goncalves-Dornelas, Higgor [2 ]
Junho Pena, Sergio Danilo [1 ,2 ]
机构
[1] Univ Fed Minas Gerais, Inst Ciencias Biol, Dept Bioquim & Imunol, BR-31270901 Belo Horizonte, MG, Brazil
[2] GENE Nucleo Genet Med, BR-30130909 Belo Horizonte, MG, Brazil
[3] Univ Sao Paulo, Inst Biociencias, Dept Genet & Biol Evolutiva, BR-05508900 Sao Paulo, Brazil
[4] PerkinElmer Inc, Signature Genom Labs, Spokane, WA 99207 USA
[5] Genet Vet Sci Inc, Paw Print Genet, Spokane, WA 99202 USA
关键词
WILLIAMS-BEUREN-SYNDROME; GENOMIC MICRODELETIONS; CLINICAL PHENOTYPE; DELETION; IDENTIFICATION; REARRANGEMENTS; BREAKPOINTS; DELINEATION; REFINEMENT; MICROARRAY;
D O I
10.1155/2014/836082
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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 10US dollars in reagent costs.
引用
收藏
页数:8
相关论文
共 50 条
  • [1] Fast, accurate, and cost-effective poultry sex genotyping using real-time polymerase chain reaction
    Cordeiro, Ciro D.
    Gonceer, Nesim
    Dorus, Steve
    Crill, James E.
    Moshayoff, Vardit
    Lachman, Amit
    Moran, Asaf
    Vilenchik, Dan
    Fedida-Metula, Shlomit
    FRONTIERS IN VETERINARY SCIENCE, 2023, 10
  • [2] Transferring a Quantitative Molecular Diagnostic Test to Multiple Real-Time Quantitative PCR Platforms
    Guertler, Claudia
    Laible, Mark
    Schwabe, Wolfgang
    Steinhaeuser, Heike
    Li, Xingmin
    Liu, Shujin
    Schlombs, Kornelia
    Sahi, Ugur
    JOURNAL OF MOLECULAR DIAGNOSTICS, 2018, 20 (04): : 398 - 414
  • [3] A cost-effective and efficient approach for generating and assembling reagents for conducting real-time PCR
    Ridim D Mote
    V Shinde Laxmikant
    Surya Bansi Singh
    Mahak Tiwari
    Hemant Singh
    Juhi Srivastava
    Vidisha Tripathi
    Vasudevan Seshadri
    Amitabha Majumdar
    Deepa Subramanyam
    Journal of Biosciences, 2021, 46
  • [4] A cost-effective and efficient approach for generating and assembling reagents for conducting real-time PCR
    Mote, Ridim D.
    Laxmikant, V. Shinde
    Singh, Surya Bansi
    Tiwari, Mahak
    Singh, Hemant
    Srivastava, Juhi
    Tripathi, Vidisha
    Seshadri, Vasudevan
    Majumdar, Amitabha
    Subramanyam, Deepa
    JOURNAL OF BIOSCIENCES, 2021, 46 (04)
  • [5] Rapid and cost-effective baculovirus sample preparation method as a viable alternative to conventional preparation for quantitative real-time PCR
    George, Steve
    Sokolenko, Stanislav
    Aucoin, Marc Gordon
    JOURNAL OF VIROLOGICAL METHODS, 2012, 182 (1-2) : 27 - 36
  • [6] Real-time PCR to detect a chromosome 1p36 deletion:: A novel screening strategy for submicroscopic subtelomeric deletions.
    Touraine, RL
    Adouard, V
    de Fréminville, B
    Pestre, S
    Till, M
    Prieur, F
    Lauras, B
    AMERICAN JOURNAL OF HUMAN GENETICS, 2001, 69 (04) : 337 - 337
  • [7] A method for accurate detection of genomic microdeletions using real-time quantitative PCR
    Rosanna Weksberg
    Simon Hughes
    Laura Moldovan
    Anne S Bassett
    Eva WC Chow
    Jeremy A Squire
    BMC Genomics, 6
  • [8] Accurate and objective copy number profiling using real-time quantitative PCR
    D'haene, Barbara
    Vandesompele, Jo
    Hellemans, Jan
    METHODS, 2010, 50 (04) : 262 - 270
  • [9] A method for accurate detection of genomic microdeletions using real-time quantitative PCR
    Weksberg, R
    Hughes, S
    Moldovan, L
    Bassett, AS
    Chow, EWC
    Squire, JA
    BMC GENOMICS, 2005, 6 (1)
  • [10] An accurate and cost-effective stereo matching algorithm and processor for real-time embedded multimedia systems
    Kyeong-ryeol Bae
    Byungin Moon
    Multimedia Tools and Applications, 2017, 76 : 17907 - 17922