Ann Lab Med.  2023 Mar;43(2):153-166. 10.3343/alm.2023.43.2.153.

Comprehensive Evaluation of the NeoBase 2 Non-derivatized MSMS Assay and Exploration of Analytes With Significantly Different Concentrations Between Term and Preterm Neonates

• Lee B
• Heo WY
• Kim JA
• Lee HS
• Hwang N
• Park HD
• Sung SI
• Chang YS
• Park WS
• Lee SY

• KMID: 2551692
• DOI: http://doi.org/10.3343/alm.2023.43.2.153

Abstract

Background
Despite the popularity of the NeoBase 2 Non-derivatized MSMS assay (PerkinElmer, Turku, Finland), there are no reports of its comprehensive evaluation, including the ability to distinguish transient tyrosinemia of the newborn (TTN) from tyrosinemia type 1 (TYR 1) using succinylacetone (SUAC). No newborn screening (NBS) cutoffs for preterm neonates in the Korean population have been suggested. We evaluated the NeoBase 2 assay and identified analytes requiring different cutoffs in preterm neonates.
Methods
Residual NBS dried blood spot samples and proficiency testing (PT) materials of the Newborn Screening Quality Assurance Program and the Korean Association of External Quality Assessment Service were used. Precision, accuracy, limit of detection (LOD), lower limit of quantification (LLOQ), linearity, recovery, carryover, and performance of SUAC were evaluated. Cutoffs were determined, and analytes requiring different cutoffs in preterm neonates were investigated.
Results
Mean CVs for within-run and between-day precision were within 15%. Accuracy analysis indicated high agreement with in-house derivatized assay results and results of other PT participants. All analytes demonstrated acceptable LOD, LLOQ, and linearity. Recoveries were acceptable, except for SUAC. Carryover was negligible. Cutoffs were established for all analytes; Tyr, adenosine, and C20:0-lysophosphatidylcholine required different cutoffs in preterm neonates. Differential diagnosis of TYR 1 and TTN was successful with simultaneous Tyr and SUAC measurement.
Conclusions
The NeoBase 2 assay demonstrated satisfactory performance. The additional analytes provide a wider diagnostic coverage, and the simultaneous measurement of Tyr and SUAC is efficient in excluding TYR 1. The new cutoffs for preterm neonates may decrease false-positive rates, without compromising diagnostic sensitivity.

Keyword

NeoBase 2 assay; Performance; Accuracy; Tandem mass spectrometry; Newborn screening; Dried blood spot; Cutoff; Preterm; Tyrosinemia; Succinylacetone

Figure

• 

  Fig. 1 Bland–Altman plots comparing the NeoBase 2 non-derivatized assay results with those of (A) NSQAP participants using the Neo-Base 2 assay for amino acids, SUAC, and acylcarnitines, (B) KEQAS participants regardless of the assay for amino acids and acylcarnitines, and (C) the in-house derivatized assay. Abbreviations: NSQAP, Newborn Screening Quality Assurance Program; SUAC, succinylacetone; KEQAS, Korean Association of External Quality Assessment Service; amino acids are abbreviated as three-letter symbols according to the International Union of Pure and Applied Chemistry (IUPAC) nomenclature.

• 

  Fig. 2 Box-and-whisker plots of selective analytes that potentially require a different cutoff in preterm neonates. Red dots and blue dots represent the values measured in preterm neonates and term neonates, respectively, and the dashed gray line indicates the established cutoff for term neonates. Abbreviations: P, Preterm; T, Term; ADO, adenosine; LPC, lysophosphatidylcholine; amino acids are abbreviated as three-letter symbols according to the International Union of Pure and Applied Chemistry (IUPAC) nomenclature.

• 

  Fig. 3 Scatter plots for (A) comparison of Tyr concentrations between the in-house derivatized assay and the NeoBase 2 assay and (B) Tyr and SUAC measured using the NeoBase 2 assay. The samples used in the analysis are composed of 42 residual clinical samples with an increased Tyr concentration and two NSQAP PT materials with an increase in both Tyr and SUAC concentrations. Abbreviations: Tyr, tyrosine; SUAC, succinylacetone; NSQAP, Newborn Screening Quality Assurance Program; PT, proficiency testing.

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