Influence of Hematocrit and Total-Spot Volume on Performance Characteristics of Dried Blood Spots for Newborn Screening

Hall, Elizabeth M.; Flores, Sharon R.; Jesús, Víctor R. De

doi:10.3390/ijns1020069

Cited by 82 publications

(74 citation statements)

References 12 publications

(9 reference statements)

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“…Despite this, however, there was still considerable inter-individual variation in the estimated volume of blood in 3 mm DBS punches from 30 different individuals using this assay (CV % 32%) (Figure 2). [8,11,23] The estimated volume of blood in a 3 mm disc obtained from a conventional newborn screening blood collection paper (903 paper) in a previous study was %3 μL, [5] which is notably higher than the estimates calculated for the 903 paper from the gravimetric analysis in the current study (2.2 AE 0.1 μL). [22] Our findings are in line with previous studies using other DBS systems that have also reported variations in Table 1.…”

Section: Resultscontrasting

confidence: 71%

“…[10][11][12]21] One such method is an on-card approach to assess the hematocrit of DBS in order to correct for sample volume, which has previously been shown to improve the precision and the accuracy of the results. As a result, several methods have been developed to account for such DBS volume variations.…”

Section: Resultsmentioning

confidence: 99%

“…[3,4] Multiple analytes can be measured from a single disc of DBS simultaneously, mainly because of the utilization of sensitive instruments such as the mass spectrometer. [7][8][9] Whereas a number of methods have been developed in an attempt to correct for these factors, including post-column infusion of an internal standard and on-card hematocrit analysis, [10][11][12] these approaches add further complexity to the procedure. These include the minimal volume of blood required (30-100 μL), ability to collect samples from a finger or heel prick, thus eliminating the need for venepuncture, ease of sample transport, and storage and stability of the compounds for months or years on the DBS once blood is dried.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Estimation of the Volume of Blood in a Small Disc Punched From a Dried Blood Spot Card

Hewawasam

Liu

Jeffery

et al. 2017

Euro J Lipid Sci & Tech

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A specialized dried blood spot (DBS) collection system (PUFAcoat™) in combination with liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) has enabled the measurement of numerous analytes in minimal volumes of blood. The current study aimed to determine the volume of blood in 3 and 6 mm discs obtained from our DBS system. The volume of blood in 3 and 6 mm discs obtained from DBS cards is estimated using four different methods: (i) gravimetric analysis; (ii) LC‐MS/MS; (iii) a hemoglobin colorimetric assay; and (iv) GC. Differences in the estimated volume are compared between methods, and variations in estimated blood volume within and between individuals are determined. The average volume of blood in a DBS disc is calculated to be 1.6 ± 0.4 μL and 8.7 ± 1.9 μL for 3 and 6 mm discs, respectively. This estimate is similar between direct and indirect analytical methods and between DBS samples with different starting volumes independent of the method, but there is considerable variation in the volume of blood in comparably‐sized DBS discs from different individuals. Current methods enable the estimation of the blood volume in a small disc obtained from a DBS but a method that can both accurately measure volume and store blood on a DBS is required. Practical Applications: This study demonstrates that it is possible to evaluate the volume of blood contained in a small disc punched from a dried blood spot (DBS) card using a range of different methods, but there is considerable variation in the estimate of volume in samples collected from different individuals. These variations could potentially result in overestimation or underestimation of endogenous levels of various metabolites if presuming a fixed volume, which may be clinically significant. Although there are several ways to correct for blood volume contained on a DBS, the practicality and the universality of these methods are questionable. Ideally, the development of a tool to better determine the volume of blood and/or to precisely spot a volume of blood is required to ensure accuracy when expressing the results of DBS analyses per unit of blood. The volume of blood in a 3 or 6 mm disc obtained from a DBS needs to be known to accurately quantify endogenous levels of metabolites per unit of blood using liquid chromatography‐tandem mass spectrometry.

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Section: Resultscontrasting

confidence: 71%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Estimation of the Volume of Blood in a Small Disc Punched From a Dried Blood Spot Card

Hewawasam

Liu

Jeffery

et al. 2017

Euro J Lipid Sci & Tech

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“…Dr. Gelb's letter also states that “there is no correlation seen in one enzymatic activity compared to another… variation in white cell count or other DBS quality factors are not dominant effects” without providing appropriate supporting data. This statement is totally incompatible with multiple peer reviewed publications to the contrary [7], [8], [9], [10]. Newborn screening for LSDs by either MS/MS or DMF results in a relatively high rate of presumptive positive results, because cut-offs are near the low limits of quantification and must be set conservatively to take into account well-documented pre-analytical variables and avoid false negatives [2].…”

mentioning

confidence: 98%

Response to Gelb et al.: “Comparison of tandem mass spectrometry to fluorimetry for newborn screening of LSDs”

Millington

2017

Molecular Genetics and Metabolism Reports

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“…Absorption time was measured with a stopwatch, which was started when the blood touched the filter paper and stopped when any shine from the liquid blood was no longer visible on the surface. Absorption diameters were measured at the widest diameter [6].…”

Section: Preparation Of Dbss On Filter Paper Treated With Gluementioning

confidence: 99%

Glue for Manufacturing Heel Prick Filter Cards Does Not Interfere with the Measurement of Analytes for Newborn Screening

Maase

Bouva

Schielen

2017

IJNS

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Population-based newborn screening (NBS) using blood collected and dried on filter paper was developed in the 1960s and remains the international standard for NBS programs. Glue, used in the manufacture of dried blood collection cards, may present a source of contamination and is often considered as a possible cause of anomalous results in routine screening. Our study evaluates this potential contamination on NBS analyses. EBF#1003 glue was blotted onto dried blood collection cards made of Whatman grade 903 filter paper (Whatman#903) and adult whole blood was pipetted onto the dried glue blots. In addition, blank glue blots (i.e., no blood) and dried blood spots (DBSs) in the absence of glue were prepared. The DBSs and blank samples were run in duplicate as routine samples for NBS. DBS absorption time and diameter, the effect of glue on measurements (concentrations and variation) were assessed. DBS absorption time and shape were equivalent for DBSs prepared in the absence and presence of undiluted glue. DBS diameter increased when prepared in the presence of glue. When EBF#1003 was diluted prior to use, DBS absorption time increased, and DBSs were non-uniform. Glue, diluted or not, did not produce measurements above the established Limit of Detection (LOD) for all assays used in the current Dutch screening programme. For all analytes with concentrations in the quantifiable range, contamination with glue had no effect on measurement variation, as it appeared equivalent to variation in untreated DBSs. Our data show that, in the unlikely event of contamination of Whatman#903 with EBF#1003, there is no effect on the measured concentration of analytes.

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Influence of Hematocrit and Total-Spot Volume on Performance Characteristics of Dried Blood Spots for Newborn Screening

Cited by 82 publications

References 12 publications

Estimation of the Volume of Blood in a Small Disc Punched From a Dried Blood Spot Card

Estimation of the Volume of Blood in a Small Disc Punched From a Dried Blood Spot Card

Response to Gelb et al.: “Comparison of tandem mass spectrometry to fluorimetry for newborn screening of LSDs”

Glue for Manufacturing Heel Prick Filter Cards Does Not Interfere with the Measurement of Analytes for Newborn Screening

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