Development of a Multiplex Real-Time PCR Assay for the Newborn Screening of SCID, SMA, and XLA

被引:27
作者
Gutierrez-Mateo, Cristina [1 ]
Timonen, Anne [2 ]
Vaahtera, Katja [2 ]
Jaakkola, Markku [2 ]
Hougaard, David M. [3 ]
Bybjerg-Grauholm, Jonas [3 ]
Baekvad-Hansen, Marie [3 ]
Adamsen, Dea [3 ]
Filippov, Galina [1 ]
Dallaire, Stephanie [1 ]
Goldfarb, David [1 ]
Schoener, Daniel [1 ]
Wu, Rongcong [1 ]
机构
[1] PerkinElmer, 940 Winter St, Waltham, MA 02451 USA
[2] Wallac Oy, PerkinElmer, Mustionkatu 6, Turku 20750, Finland
[3] Statens Serum Inst, Danish Ctr Neonatal Screening, DK-2300 Copenhagen, Denmark
关键词
Newborn Screening; SCID; SMA; XLA; DBS; real-time PCR; TREC; KREC; SMN1; SEVERE COMBINED IMMUNODEFICIENCY; SPINAL MUSCULAR-ATROPHY; DISEASES; SMN1;
D O I
10.3390/ijns5040039
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Numerous studies have shown evidence supporting the benefits of universal newborn screening for primary immunodeficiencies (PID) and for Spinal Muscular Atrophy (SMA). We have developed a four-plex, real-time PCR assay to screen for Severe Combined Immune Deficiencies (SCID), X-linked agammaglobulinemia (XLA), and SMA in DNA extracted from a single 3.2 mm punch of a dried blood spot (DBS). A simple, high-throughput, semi-automated DNA extraction method was developed for a Janus liquid handler that can process 384 DBS punches in four 96-well plates in just over one hour with sample tracking capability. The PCR assay identifies the absence of exon 7 in the SMN1 gene, while simultaneously evaluating the copy number of T-cell receptor excision circles (TREC) and Kappa-deleting recombination excision circles (KREC) molecules. Additionally, the amplification of a reference gene, RPP30, was included in the assay as a quality/quantity indicator of DNA isolated from the DBS. The assay performance was demonstrated on over 3000 DNA samples isolated from punches of putative normal newborn DBS. The reliability and analytical accuracy were further evaluated using DBS controls, and contrived and confirmed positive samples. The results from this study demonstrate the potential of future molecular DBS assays, and highlight how a multiplex assay could benefit newborn screening programs.
引用
收藏
页数:11
相关论文
共 31 条
  • [1] American College of Medical Genetics Newborn Screening Expert Group Naming and Counting Disorders (Conditions), 2006, PEDIATRICS, V117, pS308, DOI [10.1542/peds.2005-2633J, DOI 10.1542/PEDS.2005-2633J]
  • [2] American College of Medical Genetics Newborn Screening Expert Group Newborn Screening, 2006, GENET MED, V117, pS296
  • [3] Development of a routine newborn screening protocol for severe combined immunodeficiency
    Baker, Mei W.
    Grossman, William J.
    Laessig, Ronald H.
    Hoffman, Gary L.
    Brokopp, Charles D.
    Kurtycz, Daniel F.
    Cogley, Michael F.
    Litsheim, Thomas J.
    Katcher, Murray L.
    Routes, John M.
    [J]. JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, 2009, 124 (03) : 522 - 527
  • [4] Newborn Screening for Severe Primary Immunodeficiency Diseases in Sweden-a 2-Year Pilot TREC and KREC Screening Study
    Barbaro, Michela
    Ohlsson, Annika
    Borte, Stephan
    Jonsson, Susanne
    Zetterstrom, Rolf H.
    King, Jovanka
    Winiarski, Jacek
    von Dobeln, Ulrika
    Hammarstrom, Lennart
    [J]. JOURNAL OF CLINICAL IMMUNOLOGY, 2017, 37 (01) : 51 - 60
  • [5] Newborn screening for SMA in Southern Belgium
    Boemer, Francois
    Caberg, Jean-Hubert
    Dideberg, Vinciane
    Dardenne, Domien
    Bours, Vincent
    Hiligsmann, Mickael
    Dangouloff, Tamara
    Servais, Laurent
    [J]. NEUROMUSCULAR DISORDERS, 2019, 29 (05) : 343 - 349
  • [6] Combined newborn screening for familial hemophagocytic lymphohistiocytosis and severe T- and B-cell immunodeficiencies
    Borte, Stephan
    Meeths, Marie
    Liebscher, Ines
    Krist, Kerstin
    Nordenskjold, Magnus
    Hammarstrom, Lennart
    von Dobeln, Ulrika
    Henter, Jan-Inge
    Bryceson, Yenan T.
    [J]. JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, 2014, 134 (01) : 226 - 228
  • [7] Guidelines for newborn screening of primary immunodeficiency diseases
    Borte, Stephan
    von Dobelin, Ulrika
    Hammarstrom, Lennart
    [J]. CURRENT OPINION IN HEMATOLOGY, 2013, 20 (01) : 48 - 54
  • [8] Neonatal screening for severe primary immunodeficiency diseases using high-throughput triplex real-time PCR
    Borte, Stephan
    von Dobeln, Ulrika
    Fasth, Anders
    Wang, Ning
    Janzi, Magdalena
    Winiarski, Jacek
    Sack, Ulrich
    Pan-Hammarstrom, Qiang
    Borte, Michael
    Hammarstrom, Lennart
    [J]. BLOOD, 2012, 119 (11) : 2552 - 2555
  • [9] Development of population-based newborn screening for severe combined immunodeficiency
    Chan, K
    Puck, JM
    [J]. JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, 2005, 115 (02) : 391 - 398
  • [10] Presymptomatic Diagnosis of Spinal Muscular Atrophy Through Newborn Screening
    Chien, Yin-Hsiu
    Chiang, Shu-Chuan
    Weng, Wen-Chin
    Lee, Ni-Chung
    Lin, Ching-Jie
    Hsieh, Wu-Shiun
    Lee, Wang-Tso
    Jong, Yuh-Jyh
    Ko, Tsang-Ming
    Hwu, Wuh-Liang
    [J]. JOURNAL OF PEDIATRICS, 2017, 190 : 124 - +