International Journal of Neonatal Screening

Research

11 pages, 941 KiB

Open AccessArticle

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

by Cristina Gutierrez-Mateo, Anne Timonen, Katja Vaahtera, Markku Jaakkola, David M Hougaard, Jonas Bybjerg-Grauholm, Marie Baekvad-Hansen, Dea Adamsen, Galina Filippov, Stephanie Dallaire, David Goldfarb, Daniel Schoener and Rongcong Wu

Int. J. Neonatal Screen. 2019, 5(4), 39; https://doi.org/10.3390/ijns5040039 - 2 Nov 2019

Cited by 29 | Viewed by 8139

Abstract

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 [...] Read more.

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. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

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13 pages, 2757 KiB

Open AccessArticle

Factors Influencing Parental Awareness about Newborn Screening

by Věra Franková, Alena Dohnalová, Karolína Pešková, Renata Hermánková, Riona O’Driscoll, Pavel Ješina and Viktor Kožich

Int. J. Neonatal Screen. 2019, 5(3), 35; https://doi.org/10.3390/ijns5030035 - 18 Sep 2019

Cited by 13 | Viewed by 4722

Abstract

Appropriate and timely education about newborn screening (NBS) helps to foster benefits such as prompt follow up, to promote parents’ autonomy via informed consent and minimize the harms such as reducing the impact of NBS false-positive results. The aim of this study was [...] Read more.

Appropriate and timely education about newborn screening (NBS) helps to foster benefits such as prompt follow up, to promote parents’ autonomy via informed consent and minimize the harms such as reducing the impact of NBS false-positive results. The aim of this study was to ascertain how mothers are informed about NBS in the Czech Republic and to identify the variables associated with awareness about NBS. The questionnaires evaluating awareness and its determinants were mailed to a random sample of 3000 mothers 3 months post-delivery. The overall response rate was 42%. We analysed 1100 questionnaires and observed that better awareness about NBS was significantly associated with age, parity, number of information sources, child health status, size of maternity hospital and an obstetrician as the source of prenatally obtained information. Although the majority of mothers (77%) in our study recalled being informed by a physician or nurse in the neonatal ward, results have revealed that over 40% of participants did not have sufficient awareness about the principal aspects of NBS. Several measures including seminars for healthcare providers and the development and distribution of new educational materials were adopted to improve parental education about NBS in the Czech Republic. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

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15 pages, 1477 KiB

Open AccessArticle

Wisconsin’s Screening Algorithm for the Identification of Newborns with Congenital Adrenal Hyperplasia

by Eric R. Bialk, Michael R. Lasarev and Patrice K. Held

Int. J. Neonatal Screen. 2019, 5(3), 33; https://doi.org/10.3390/ijns5030033 - 6 Sep 2019

Cited by 18 | Viewed by 6252

Abstract

Newborn screening for congenital adrenal hyperplasia (CAH) has one of the highest false positive rates of any of the diseases on the Wisconsin panel. This is largely due to the first-tier immune assay cross-reactivity and physiological changes in the concentration of 17-hydroxyprogesterone during [...] Read more.

Newborn screening for congenital adrenal hyperplasia (CAH) has one of the highest false positive rates of any of the diseases on the Wisconsin panel. This is largely due to the first-tier immune assay cross-reactivity and physiological changes in the concentration of 17-hydroxyprogesterone during the first few days of life. To improve screening for CAH, Wisconsin developed a second-tier assay to quantify four different steroids (17-hydroxyprogesterone, 21-deoxycortisol, androstenedione, and cortisol) by liquid chromatography–tandem mass spectrometry (LC–MSMS) in dried blood spots. From validation studies which included the testing of confirmed CAH patients, Wisconsin established its own reporting algorithm that incorporates steroid concentrations as well as two different ratios—the birth weight and the collection time—to identify babies at risk for CAH. Using the newly developed method and algorithm, the false positive rate for the CAH screening was reduced by 95%. Patients with both classical forms of CAH, salt-wasting and simple virilizing, were identified. This study replicates and expands upon previous work to develop a second-tier LC–MSMS steroid profiling screening assay for CAH. The validation and prospective study results provide evidence for an extensive reporting algorithm that incorporates multiple steroids, birth weight, and collection times. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

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12 pages, 716 KiB

Open AccessArticle

Initial Evaluation of Prospective and Parallel Assessments of Cystic Fibrosis Newborn Screening Protocols in Eastern Andalusia: IRT/IRT versus IRT/PAP/IRT

by Ilham Sadik, Inmaculada Pérez de Algaba, Rocío Jiménez, Carmen Benito, Javier Blasco-Alonso, Pilar Caro, Víctor M. Navas-López, Javier Pérez-Frías, Estela Pérez, Juliana Serrano and Raquel Yahyaoui

Int. J. Neonatal Screen. 2019, 5(3), 32; https://doi.org/10.3390/ijns5030032 - 3 Sep 2019

Cited by 4 | Viewed by 3644

Abstract

Identifying newborns at risk for cystic fibrosis (CF) by newborn screening (NBS) using dried blood spot (DBS) specimens provides an opportunity for presymptomatic detection. All NBS strategies for CF begin with measuring immunoreactive trypsinogen (IRT). Pancreatitis-associated protein (PAP) has been suggested as second-tier [...] Read more.

Identifying newborns at risk for cystic fibrosis (CF) by newborn screening (NBS) using dried blood spot (DBS) specimens provides an opportunity for presymptomatic detection. All NBS strategies for CF begin with measuring immunoreactive trypsinogen (IRT). Pancreatitis-associated protein (PAP) has been suggested as second-tier testing. The main objective of this study was to evaluate the analytical performance of an IRT/PAP/IRT strategy versus the current IRT/IRT strategy over a two-year pilot study including 68,502 newborns. The design of the study, carried out in a prospective and parallel manner, allowed us to compare four different CF-NBS protocols after performing a post hoc analysis. The best PAP cutoff point and the potential sources of PAP false positive results in our non-CF newborn population were also studied. 14 CF newborns were detected, resulting in an overall CF prevalence of 1/4, 893 newborns. The IRT/IRT algorithm detected all CF cases, but the IRT/PAP/IRT algorithm failed to detect one case of CF. The IRT/PAP/IRT with an IRT-dependent safety net protocol was a good alternative to improve sensitivity to 100%. The IRT × PAP/IRT strategy clearly performed better, with a sensitivity of 100% and a positive predictive value (PPV) of 39%. Our calculated optimal cutoffs were 2.31 µg/L for PAP and 167.4 µg²/L² for IRT × PAP. PAP levels were higher in females and newborns with low birth weight. PAP false positive results were found mainly in newborns with conditions such as prematurity, sepsis, and hypoxic-ischemic encephalopathy. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

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9 pages, 861 KiB

Open AccessArticle

Newborn Sickle Cell and Thalassaemia Screening Programme: Automating and Enhancing the System to Evaluate the Screening Programme

by Catherine Coppinger and Robyn O’Loughlin

Int. J. Neonatal Screen. 2019, 5(3), 30; https://doi.org/10.3390/ijns5030030 - 31 Aug 2019

Cited by 2 | Viewed by 2978

Abstract

Good information is needed to demonstrate that a screening programme is meeting its objectives, to measure performance against standards and to ensure that action is taken if standards are not met. In 2010, the NHS Sickle Cell and Thalassaemia (SCT) Screening Programme established [...] Read more.

Good information is needed to demonstrate that a screening programme is meeting its objectives, to measure performance against standards and to ensure that action is taken if standards are not met. In 2010, the NHS Sickle Cell and Thalassaemia (SCT) Screening Programme established a process to collect data on the main outcome measures for newborn babies. In 2016, a review identified that data completeness and quality relied on manual processes and there was widespread dissatisfaction amongst data providers due to duplication of data entry, poor feedback and lack of oversight of the baby to ensure safe handover from screening into treatment services. Using an Agile service design process and following the Government Digital Service Model, the SCT Screening Programme worked in close collaboration with users, wider stakeholders and system suppliers to design and build a new automated system. The new system ensures that the screening programme can fulfil its duty to evaluate the effectiveness of the programme, whilst pleasing the users and enhancing safety. User experience must be central to design and ongoing development to ensure that a new IT system is fit for purpose and adopted by users. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

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5 pages, 185 KiB

Open AccessArticle

Diagnosis of Carnitine Deficiency in Extremely Preterm Neonates Related to Parenteral Nutrition: Two Step Newborn Screening Approach

by Mamatha Ramaswamy, Victor Anthony Skrinska, Rola Fayez Mitri and Ghassan Abdoh

Int. J. Neonatal Screen. 2019, 5(3), 29; https://doi.org/10.3390/ijns5030029 - 31 Aug 2019

Cited by 7 | Viewed by 3074

Abstract

Currently, there is no evidence in the literature to support the routine supplementation of all parenterally fed premature infants with l-carnitine. In our study, we found that about 8.56% of extremely preterm neonates are diagnosed with carnitine deficiency secondary to malnutrition, either [...] Read more.

Currently, there is no evidence in the literature to support the routine supplementation of all parenterally fed premature infants with l-carnitine. In our study, we found that about 8.56% of extremely preterm neonates are diagnosed with carnitine deficiency secondary to malnutrition, either due to reduced stores at birth or related to total parenteral nutrition (TPN). Our two step approach of performing newborn screening (NBS) again at 32 weeks gestational age (GA) equivalent helps to diagnose 81.4% more preterm babies with carnitine deficiency—who would otherwise be missed—and supplement them with l-carnitine for optimal growth. We performed a retrospective cohort study to diagnose carnitine deficiency related to malnutrition in two groups: those presenting at birth and those presenting later in life. We found that there was a statistically significant difference in the median GA and birth weight (BW) between the two groups, but there was no difference in the free carnitine levels. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

15 pages, 1244 KiB

Open AccessArticle

A Cost-Effectiveness Analysis of Newborn Screening for Severe Combined Immunodeficiency in the UK

by Alice Bessey, James Chilcott, Joanna Leaviss, Carmen de la Cruz and Ruth Wong

Int. J. Neonatal Screen. 2019, 5(3), 28; https://doi.org/10.3390/ijns5030028 - 30 Aug 2019

Cited by 17 | Viewed by 5372

Abstract

Severe combined immunodeficiency (SCID) can be detected through newborn bloodspot screening. In the UK, the National Screening Committee (NSC) requires screening programmes to be cost-effective at standard UK thresholds. To assess the cost-effectiveness of SCID screening for the NSC, a decision-tree model with [...] Read more.

Severe combined immunodeficiency (SCID) can be detected through newborn bloodspot screening. In the UK, the National Screening Committee (NSC) requires screening programmes to be cost-effective at standard UK thresholds. To assess the cost-effectiveness of SCID screening for the NSC, a decision-tree model with lifetable estimates of outcomes was built. Model structure and parameterisation were informed by systematic review and expert clinical judgment. A public service perspective was used and lifetime costs and quality-adjusted life years (QALYs) were discounted at 3.5%. Probabilistic, one-way sensitivity analyses and an exploratory disbenefit analysis for the identification of non-SCID patients were conducted. Screening for SCID was estimated to result in an incremental cost-effectiveness ratio (ICER) of £18,222 with a reduction in SCID mortality from 8.1 (5–12) to 1.7 (0.6–4.0) cases per year of screening. Results were sensitive to a number of parameters, including the cost of the screening test, the incidence of SCID and the disbenefit to the healthy at birth and false-positive cases. Screening for SCID is likely to be cost-effective at £20,000 per QALY, key uncertainties relate to the impact on false positives and the impact on the identification of children with non-SCID T Cell lymphopenia. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

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11 pages, 1733 KiB

Open AccessArticle

Duchenne Muscular Dystrophy Newborn Screening: Evaluation of a New GSP^® Neonatal Creatine Kinase-MM Kit in a US and Danish Population

by Anne Timonen, Michele Lloyd-Puryear, David M. Hougaard, Liisa Meriö, Pauliina Mäkinen, Ville Laitala, Tuukka Pölönen, Kristin Skogstrand, Annie Kennedy, Sari Airenne, Hanna Polari and Teemu Korpimäki

Int. J. Neonatal Screen. 2019, 5(3), 27; https://doi.org/10.3390/ijns5030027 - 27 Aug 2019

Cited by 30 | Viewed by 7418

Abstract

Duchenne muscular dystrophy (DMD/Duchenne) is a progressive X-linked disease and is the most common pediatric-onset form of muscular dystrophy, affecting approximately 1:5000 live male births. DNA testing for mutations in the dystrophin gene confirms the diagnosis of this disorder. This study involves assessment [...] Read more.

Duchenne muscular dystrophy (DMD/Duchenne) is a progressive X-linked disease and is the most common pediatric-onset form of muscular dystrophy, affecting approximately 1:5000 live male births. DNA testing for mutations in the dystrophin gene confirms the diagnosis of this disorder. This study involves assessment of screening newborns for DMD using an immunoassay for muscle-type (MM) creatine kinase (CK) isoform—the GSP Neonatal CK-MM kit. Comparisons were made with CK activity determination by fluorescence measurement. In addition, the study evaluated the effect of gestational age, age of infant at time of sampling and how stable the CK-MM was over time. This assay discriminates well between normal, unaffected and Duchenne affected populations and is suitable for Duchenne newborn screening. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

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9 pages, 1549 KiB

Open AccessArticle

Validation of a Fast, Robust, Inexpensive, Two-Tiered Neonatal Screening Test algorithm on Dried Blood Spots for Spinal Muscular Atrophy

by Annuska Strunk, Andre Abbes, Antoine R. Stuitje, Chris Hettinga, Eline M. Sepers, Reinier Snetselaar, Jan Schouten, Fay-Lynn Asselman, Inge Cuppen, Henny Lemmink, W. Ludo van der Pol and Henk Engel

Int. J. Neonatal Screen. 2019, 5(2), 21; https://doi.org/10.3390/ijns5020021 - 15 May 2019

Cited by 13 | Viewed by 5980

Abstract

Spinal muscular atrophy (SMA) is one of the leading genetic causes of infant mortality with an incidence of 1:10,000. The recently-introduced antisense oligonucleotide treatment improves the outcome of this disease, in particular when applied at an early stage of progression. The genetic cause [...] Read more.

Spinal muscular atrophy (SMA) is one of the leading genetic causes of infant mortality with an incidence of 1:10,000. The recently-introduced antisense oligonucleotide treatment improves the outcome of this disease, in particular when applied at an early stage of progression. The genetic cause of SMA is, in >95% of cases, a homozygous deletion of the survival motor neuron 1 (SMN1) gene, which makes the low-cost detection of SMA cases as part of newborn screening programs feasible. We developed and validated a new SALSA MC002 melting curve assay that detects the absence of the SMN1 exon 7 DNA sequence without detecting asymptomatic carriers and reliably discriminates SMN1 from its genetic homolog SMN2 using crude extracts from newborn screening cards. Melting curve analysis shows peaks specific for both the SMN1 gene and the disease modifying SMN2 homolog. The detection of the SMN2 homolog, of which the only clinically relevant difference from the SMN1 gene is a single nucleotide in exon 7, was only used to confirm a correct reaction in samples that lacked the SMN1 gene, and not for SMN2 quantification. We retrieved 47 DBS samples from children with genetically-confirmed SMA, after informed consent from parents, and 375 controls from the national archive of the Dutch National Institute for Public Health and the Environment (RIVM). The assay correctly identified all anonymized and randomized SMA and control samples (i.e., sensitivity and specificity of 100%), without the detection of carriers, on the three most commonly-used PCR platforms with melting curve analysis. This test’s concordance with the second-tier ‘golden standard’ P021 SMA MLPA test was 100%. Using the new P021–B1 version, crude extracts from DBS cards could also be used to determine the SMN2 copy number of SMA patients with a high level of accuracy. The MC002 test showed the feasibility and accuracy of SMA screening in a neonatal screening program. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

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11 pages, 779 KiB

Open AccessArticle

Including Classical Galactosaemia in the Expanded Newborn Screening Panel Using Tandem Mass Spectrometry for Galactose-1-Phosphate

by Arieh S. Cohen, Marta Baurek, Allan M. Lund, Morten Dunø and David M. Hougaard

Int. J. Neonatal Screen. 2019, 5(2), 19; https://doi.org/10.3390/ijns5020019 - 4 May 2019

Cited by 9 | Viewed by 5714

Abstract

Galactosaemia has been included in various newborn screening programs since 1963. Several methods are used for screening; however, the predominant methods used today are based on the determination of either galactose-1-phosphate uridyltransferase (GALT) activity or the concentration of total galactose. These methods cannot [...] Read more.

Galactosaemia has been included in various newborn screening programs since 1963. Several methods are used for screening; however, the predominant methods used today are based on the determination of either galactose-1-phosphate uridyltransferase (GALT) activity or the concentration of total galactose. These methods cannot be multiplexed and therefore require one full punch per sample. Since the introduction of mass spectrometry in newborn screening, many diseases have been included in newborn screening programs. Here, we present a method for including classical galactosaemia in an expanded newborn screening panel based on the specific determination of galactose-1-phosphate by tandem mass spectrometry. The existing workflow only needs minor adjustments, and it can be run on the tandem mass spectrometers in routine use. Furthermore, compared to the currently used methods, this novel method has a superior screening performance, producing significantly fewer false positive results. We present data from 5500 routine newborn screening samples from the Danish Neonatal Screening Biobank. The cohort was enriched by including 14 confirmed galactosaemia positive samples and 10 samples positive for other metabolic disorders diagnosed through the Danish newborn screening program. All galactosaemia positive samples were identified by the method with no false positives. Furthermore, the screening performance for other metabolic disorders was unaffected. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

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12 pages, 739 KiB

Open AccessArticle

The Importance of Assay Imprecision near the Screen Cutoff for Newborn Screening of Lysosomal Storage Diseases

by Bruce H. Robinson and Michael H. Gelb

Int. J. Neonatal Screen. 2019, 5(2), 17; https://doi.org/10.3390/ijns5020017 - 27 Mar 2019

Cited by 8 | Viewed by 3786

Abstract

For newborn screening (NBS) of lysosomal storage diseases, programs measure enzymatic activities in dried blood spots (DBS) and, in most cases, act on samples where the measurement is below a specific cutoff value. The rate of false positives and negatives in any NBS [...] Read more.

For newborn screening (NBS) of lysosomal storage diseases, programs measure enzymatic activities in dried blood spots (DBS) and, in most cases, act on samples where the measurement is below a specific cutoff value. The rate of false positives and negatives in any NBS program is of critical importance. The measured values across a population of newborns are governed by many factors, and in this article we focus on assay imprecision. Assay parameters including the Analytical Range and the Z-Factor have been discussed as a way to compare assay performance for NBS of lysosomal storage diseases. Here we show that these parameters are not rigorously connected to the rate of false positives and negatives. Rather, it is the assay imprecision near the screen cutoff that is the most important parameter that determines the rate of false positives and negatives. We develop the theoretical treatment of assay imprecision and how it is linked to screen performance. What emerges is a useful type of parametric plot that allows for rigorous assessment of the effect of assay imprecision on the rate of false positives and false negatives that is independent of the choice of screen cutoff value. Such plots are useful in choosing cutoff values. They also show that a high assay imprecision cannot be overcome by changing the cutoff value or by use of postanalysis, statistical tools. Given the importance of assay imprecision near the cutoff, we propose that quality control DBS are most useful if they span a range of analyte values near the cutoff. Our treatment is also appropriate for comparing the performance of multiple assay platforms that each measure the same quantity (i.e., the enzymatic activity in DBS). The analysis shows that it is always best to use the assay platform that gives the lowest imprecision near the cutoff. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

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Review

Jump to: Research, Other

9 pages, 1444 KiB

Open AccessReview

Newborn Screening: Current Status in Alberta, Canada

by Andy De Souza, Vanessa Wolan, Angie Battochio, Susan Christian, Stacey Hume, Grace Johner, Margaret Lilley, Ross Ridsdale, Kareena Schnabl, Chi Tran, Jolene Yuen-Jung and Iveta Sosova

Int. J. Neonatal Screen. 2019, 5(4), 37; https://doi.org/10.3390/ijns5040037 - 1 Oct 2019

Cited by 11 | Viewed by 6372

Abstract

Newborn screening (NBS) in Alberta is delivered by a number of government and health service entities who work together to provide newborn screening to infants born in Alberta, the Northwest Territories, and the Kitikmeot region of the Nunavut territory. The Alberta panel screens [...] Read more.

Newborn screening (NBS) in Alberta is delivered by a number of government and health service entities who work together to provide newborn screening to infants born in Alberta, the Northwest Territories, and the Kitikmeot region of the Nunavut territory. The Alberta panel screens for 21 disorders (16 metabolic, two endocrine, cystic fibrosis, severe combined immunodeficiency, and sickle cell disease). NBS is a standard of care, but is not mandatory. NBS performance is monitored by the Alberta Newborn Metabolic Screening (NMS) Program and NMS Laboratory, who strive for continuous quality improvement. Performance analysis found that over 99% of registered infants in Alberta received a newborn screen and over 98% of these infants received a screen result within 10 days of age. Full article

(This article belongs to the Special Issue Selected Papers from 11th ISNS European Regional Meeting)

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