Newborn Sickle Cell Disease Screening Using Electrospray Tandem Mass Spectrometry

Daniel, Yvonne; Turner, Charles

doi:10.3390/ijns4040035

Cited by 6 publications

(7 citation statements)

References 10 publications

(18 reference statements)

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“…The data obtained by this work are in agreement with other studies in the literature − and can be used to support the hypotheses addressed and the promising applicability of the validated method in both the diagnosis and control of diabetes and in the monitoring of available methods. This challenge analyte quantification was chosen to demonstrate the full capability of HRMS to differentiate high molecular weight molecules, in complex matrices with minimal sample preparation.…”

Section: Resultssupporting

confidence: 89%

New Validated Method for Quantification of Glycated Hemoglobin by LC-QToF-MS: Is HRMS Able to Quantify Clinical Samples?

Cerqueira

Fachi

Kawagushi

et al. 2020

J. Am. Soc. Mass Spectrom.

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High-resolution mass spectrometry is a powerful tool in clinical analysis but remains less explored due to its lower dynamic range and sensitivity compared to triple quadrupoles. Glycated hemoglobin (HbA1c) is the current gold standard biomarker to monitor the control of diabetes, representing long-term plasma glycemic levels. Due to its clinical importance, several methods have been developed for HbA1c quantification, using different principles; however, the results obtained with these techniques may differ according to the method adopted. Hence, there is a great need to standardize the current methods to quantify glycated hemoglobin. A new UPLC-QToF-MS method was fully validated and tested to quantify HbA1c in human samples. The peptides VHLTPE m/z 695.373 and gly-VHLTPE m/z 857.426, obtained via Glu-C digestion, were the selected peptides for quantification of HbA1c (mmol/mol). Chromatographic separation was obtained in a C18 column, maintained at 40 °C. The mobile phase was composed of water and acetonitrile, both containing 0.02% TFA and 0.1% acetic acid, and eluted in gradient mode. The method was fully validated, being considered linear in the range of 25−107 mmol/mol of HbA1c, and was sensitive, selective, precise, accurate, and free of matrix and carryover effects. The method was successfully applied to real samples, reaching about 90% agreement with reference method results, providing accurate and precise information on peptide mass, without laborious sample preparation. These results support the use of HRMS to improve the quality of quantitative results of HbA1c in health services and demonstrate a possible application of peptide investigation for clinical analysis in the near future.

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Section: Resultssupporting

confidence: 89%

New Validated Method for Quantification of Glycated Hemoglobin by LC-QToF-MS: Is HRMS Able to Quantify Clinical Samples?

Cerqueira

Fachi

Kawagushi

et al. 2020

J. Am. Soc. Mass Spectrom.

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“…The screening programme will also detect certain other variant haemoglobins by virtue of the analytical methods currently used. Additionally, the finding of Hb F only or of a very low percentage of haemoglobin A (1%–5% or less of the total haemoglobin or above the mass spectrometry action value) 8 on the newborn screen will identify the majority of babies with β thalassaemia major/TDT, necessitating follow‐up and further testing.…”

Section: Methodsmentioning

confidence: 99%

Significant haemoglobinopathies: A guideline for screening and diagnosis

Bain

Daniel

Henthorn³

et al. 2023

Br J Haematol

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Disorders of globin chain synthesis, both thalassaemia and haemoglobin variants, are common in the United Kingdom and constitute a significant public health problem. Diagnosis may be required: (i) to confirm a provisional clinical diagnosis, such as sickle cell disease (SCD) or β thalassaemia major (now often referred to as transfusion-dependent thalassaemia or TDT); (ii) to explain a haematological abnormality, such as anaemia or microcytosis; (iii) to identify an abnormality in the presymptomatic phase, as in neonatal screening; (iv) to identify foetuses at risk of significant haemoglobinopathies and offer the parents informed choice; (v) to permit genetic counselling of prospective parents; (vi) to identify the presence of sickle cell haemoglobin preoperatively. Improved fully automated systems and reagents for techniques such as cation-exchange high-performance liquid chromatography (HPLC) and capillary electrophoresis

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“…A commercial MS/MS-based hemoglobinopathy screening kit was selected as the method for differentiation because the analytical platform already existed in our laboratory and there were published reports on, e.g., the diagnostic sensitivity and specificity achieved with the kit [21,32]. As part of the implementation of the assay, tuning experiments were performed to evaluate manufacturer-recommended and previously published mass transitions for the target peptides within the scope of the kit.…”

Section: Tandem-ms Analysesmentioning

confidence: 99%

“…The SpotOn MS/MS assay for SCD screening is originally designed as a 1 st -tier analysis and its application has been previously reported by multiple groups [21,32]. Here, it has been used as a 2 nd -tier assay which required the preparation of aliquots from the original kit reagents.…”

Section: Plos Onementioning

confidence: 99%

A high-throughput newborn screening approach for SCID, SMA, and SCD combining multiplex qPCR and tandem mass spectrometry

Tesorero

Janda²,

Hörster³

et al. 2023

PLoS ONE

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Early diagnosis of severe combined immunodeficiency (SCID), spinal muscular atrophy (SMA), and sickle cell disease (SCD) improves health outcomes by providing a specific treatment before the onset of symptoms. A high-throughput nucleic acid-based method in newborn screening (NBS) has been shown to be fast and cost-effective in the early detection of these diseases. Screening for SCD has been included in Germany’s NBS Program since Fall 2021 and typically requires high-throughput NBS laboratories to adopt analytical platforms that are demanding in terms of instrumentation and personnel. Thus, we developed a combined approach applying a multiplexed quantitative real-time PCR (qPCR) assay for simultaneous SCID, SMA, and 1st-tier SCD screening, followed by a tandem mass spectrometry (MS/MS) assay for 2nd-tier SCD screening. DNA is extracted from a 3.2-mm dried blood spot from which we simultaneously quantify T-cell receptor excision circles for SCID screening, identify the homozygous SMN1 exon 7 deletion for SMA screening, and determine the integrity of the DNA extraction through the quantification of a housekeeping gene. In our two-tier SCD screening strategy, our multiplex qPCR identifies samples carrying the HBB: c.20A>T allele that is coding for sickle cell hemoglobin (HbS). Subsequently, the 2nd tier MS/MS assay is used to distinguish heterozygous HbS/A carriers from samples of patients with homozygous or compound heterozygous SCD. Between July 2021 and March 2022, 96,015 samples were screened by applying the newly implemented assay. The screening revealed two positive SCID cases, while 14 newborns with SMA were detected. Concurrently, the qPCR assay registered HbS in 431 samples which were submitted to 2nd-tier SCD screening, resulting in 17 HbS/S, five HbS/C, and two HbS/β thalassemia patients. The results of our quadruplex qPCR assay demonstrate a cost-effective and fast approach for a combined screening of three diseases that benefit from nucleic-acid based methods in high-throughput NBS laboratories.

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Newborn Sickle Cell Disease Screening Using Electrospray Tandem Mass Spectrometry

Cited by 6 publications

References 10 publications

New Validated Method for Quantification of Glycated Hemoglobin by LC-QToF-MS: Is HRMS Able to Quantify Clinical Samples?

New Validated Method for Quantification of Glycated Hemoglobin by LC-QToF-MS: Is HRMS Able to Quantify Clinical Samples?

Significant haemoglobinopathies: A guideline for screening and diagnosis

A high-throughput newborn screening approach for SCID, SMA, and SCD combining multiplex qPCR and tandem mass spectrometry

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