Newborn Screening For Severe Combined Immunodeficiency In Iowa: TREC Assay Results and Characteristics Of SCID and T-Cell Lymphopenic Patients

2014 ◽  
Vol 133 (2) ◽  
pp. AB94 ◽  
Author(s):  
Farheen Mirza ◽  
Emily Phillips ◽  
Travis Henry ◽  
Mary Beth Fasano
Keyword(s):  
T Cell ◽  
Newborn Screening ◽  
Severe Combined Immunodeficiency ◽  
Combined Immunodeficiency ◽  
Trec Assay

scholarly journals A Multiplex Immunoassay Using the Guthrie Specimen to Detect T-Cell Deficiencies Including Severe Combined Immunodeficiency Disease

2010 ◽  
Vol 56 (9) ◽  
pp. 1460-1465 ◽  
Author(s):  
David K Janik ◽  
Barbara Lindau-Shepard ◽  
Anne Marie Comeau ◽  
Kenneth A Pass
Keyword(s):  
T Cell ◽  
Newborn Screening ◽  
Clinical Status ◽  
Severe Combined Immunodeficiency ◽  
Cross Reactivity ◽  
Multiplex Assay ◽  
Molecular Technique ◽  
Combined Immunodeficiency ◽  
Pilot Studies ◽  
Trec Assay

BACKGROUND Severe combined immunodeficiency (SCID) fulfills many of the requirements for addition to a newborn screening panel. Two newborn screening SCID pilot studies are now underway using the T-cell receptor excision circle (TREC) assay, a molecular technique. Here we describe an immunoassay with CD3 as a marker for T cells and CD45 as a marker for total leukocytes that can be used with the Guthrie specimen. METHODS The multiplexing capabilities of the Luminex platform were used. Antibody pairs were used to capture and detect CD3 and CD45 from a single 3-mm punch of the Guthrie specimen. The assay for each biomarker was developed separately in identical buffers and then combined to create a multiplex assay. RESULTS Using calibrators made from known amounts of leukocytes, a detection limit of 0.25 × 106 cells/mL for CD3 and 0.125 × 106 cells/mL for CD45 was obtained. Affinity tests showed no cross-reactivity between the antibodies to CD3 and CD45. The multiplex assay was validated against 8 coded specimens of known clinical status and linked to results from the TREC assay that had identified them. All were correctly identified by the CD345 assay. CONCLUSIONS The performance parameters of the CD345 assay met the performance characteristics generally accepted for immunoassays. Our assay classifications of positive specimens concur with previous TREC results. This CD345 assay warrants evaluation as a viable alternative or complement to the TREC assay as a primary screening tool for detecting T-cell immunodeficiencies, including SCID, in Guthrie specimens.

scholarly journals Newborn Screening for Severe Combined Immunodeficiency Using the Multiple of the Median Values of T-Cell Receptor Excision Circles

2021 ◽  
Vol 7 (3) ◽  
pp. 43
Author(s):  
Michael F. Cogley ◽  
Amy E. Wiberley-Bradford ◽  
Sean T. Mochal ◽  
Sandra J. Dawe ◽  
Zachary D. Piro ◽  
...  
Keyword(s):  
T Cell ◽  
Newborn Screening ◽  
T Cell Receptor ◽  
Severe Combined Immunodeficiency ◽  
Cell Receptor ◽  
Combined Immunodeficiency ◽  
Individual Test ◽  
Copy Numbers ◽  
Excision Circles ◽  
Trec Assay

All newborn screening programs screen for severe combined immunodeficiency by measurement of T-cell receptor excision circles (TRECs). Herein, we report our experience of reporting TREC assay results as multiple of the median (MoM) rather than using conventional copy numbers. This modification simplifies the assay by eliminating the need for standards with known TREC copy numbers. Furthermore, since MoM is a measure of how far an individual test result deviates from the median, it allows normalization of TREC assay data from different laboratories, so that individual test results can be compared regardless of the particular method, assay, or reagents used.

Newborn screening for severe combined immunodeficiency using a novel and simplified method to measure T-cell excision circles (TREC)

2017 ◽  
Vol 175 ◽  
pp. 51-55 ◽  
Author(s):  
Laura Tagliaferri ◽  
Joachim B. Kunz ◽  
Margit Happich ◽  
Susanna Esposito ◽  
Thomas Bruckner ◽  
...  
Keyword(s):  
T Cell ◽  
Newborn Screening ◽  
Severe Combined Immunodeficiency ◽  
Combined Immunodeficiency ◽  
Simplified Method ◽  
Excision Circles

scholarly journals Newborn Blood Spot Screening Test Using Multiplexed Real-Time PCR to Simultaneously Screen for Spinal Muscular Atrophy and Severe Combined Immunodeficiency

2015 ◽  
Vol 61 (2) ◽  
pp. 412-419 ◽  
Author(s):  
Jennifer L Taylor ◽  
Francis K Lee ◽  
Golriz Khadem Yazdanpanah ◽  
John F Staropoli ◽  
Mei Liu ◽  
...  
Keyword(s):  
T Cell ◽  
Motor Neuron ◽  
Real Time ◽  
Real Time Pcr ◽  
Muscular Atrophy ◽  
Severe Combined Immunodeficiency ◽  
Combined Immunodeficiency ◽  
Smn1 Gene ◽  
Trec Assay ◽  
Blood Spot

Abstract BACKGROUND Spinal muscular atrophy (SMA) is a motor neuron disorder caused by the absence of a functional survival of motor neuron 1, telomeric (SMN1) gene. Type I SMA, a lethal disease of infancy, accounts for the majority of cases. Newborn blood spot screening (NBS) to detect severe combined immunodeficiency (SCID) has been implemented in public health laboratories in the last 5 years. SCID detection is based on real-time PCR assays to measure T-cell receptor excision circles (TREC), a byproduct of T-cell development. We modified a multiplexed real-time PCR TREC assay to simultaneously determine the presence or absence of the SMN1 gene from a dried blood spot (DBS) punch in a single reaction well. METHOD An SMN1 assay using a locked nucleic acid probe was initially developed with cell culture and umbilical cord blood (UCB) DNA extracts, and then integrated into the TREC assay. DBS punches were placed in 96-well arrays, washed, and amplified directly using reagents specific for TREC, a reference gene [ribonuclease P/MRP 30kDa subunit (RPP30)], and the SMN1 gene. The assay was tested on DBS made from UCB units and from peripheral blood samples of SMA-affected individuals and their family members. RESULTS DBS made from SMA-affected individuals showed no SMN1-specific amplification, whereas DBS made from all unaffected carriers and UCB showed SMN1 amplification above a well-defined threshold. TREC and RPP30 content in all DBS were within the age-adjusted expected range. CONCLUSIONS SMA caused by the absence of SMN1 can be detected from the same DBS punch used to screen newborns for SCID.

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