Rapid, single-step assay for Hunter syndrome in dried blood spots using digital microfluidics

Sista, Ramakrishna S.; Eckhardt, Allen E.; Wang, Tong; Sellos-Moura, M.; Pamula, Vamsee K.

doi:10.1016/j.cca.2011.06.015

Cited by 26 publications

(29 citation statements)

References 9 publications

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“…However, there is no basis for the statement at this time that the fluorometric assay is faster. After a 16-h incubation, we achieved an activity for the ID2S enzyme that was approximately 100-fold above the signal measured in the no-blood control, whereas the increase was approximately 20-fold in the fluorometric assay carried out with an incubation time of 1 h (18). Thus, it appears that the 2 assays have comparable sensitivities and that a few hours’ incubation time is also achievable with the current LC-MS/MS assay.…”

Section: Discussionmentioning

confidence: 92%

“…Fluorometric methods are also being developed for newborn screening of LSDs (17, 18). It is possible that both MS/MS and fluorometric methods will be implemented in newborn screening laboratories for LSDs.…”

Section: Discussionmentioning

confidence: 99%

“…It is possible that both MS/MS and fluorometric methods will be implemented in newborn screening laboratories for LSDs. One study claimed that the fluorometric method was more rapid than the MS/MS method for assaying ID2S, since the fluorometric method used a 1-h incubation and the MS/MS method used an overnight incubation (18). However, there is no basis for the statement at this time that the fluorometric assay is faster.…”

Section: Discussionmentioning

confidence: 99%

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High-Throughput Assay of 9 Lysosomal Enzymes for Newborn Screening

Spáčil¹,

Tatipaka²,

Barcenas³

et al. 2013

Clinical Chemistry

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BACKGROUND There is interest in newborn screening of lysosomal storage diseases (LSDs) because of the availability of treatments. Pilot studies have used tandem mass spectrometry with flow injection of samples to achieve multiplex detection of enzyme products. We report a multiplexing method of 9 enzymatic assays that uses HPLC-tandem mass spectrometry (MS/MS). METHODS The assay of 9 enzymes was carried out in 1 or 2 buffers with a cassette of substrates and internal standards and 1 or 2 punches of a dried blood spot (DBS) from a newborn screening card as the source of enzymes. The pre–HPLC-MS/MS sample preparation required only 4 liquid transfers before injection into a dual-column HPLC equipped with switching valves to direct the flow to separation and column equilibration. Product-specific and internal standard–specific ion fragmentations were used for MS/MS quantification in the selected reaction monitoring mode. RESULTS Analysis of blood spots from 58 random newborns and lysosomal storage disease–affected patients showed that the assay readily distinguished affected from nonaffected individuals. The time per 9-plex analysis (1.8 min) was sufficiently short to be compatible with the workflow of newborn screening laboratories. CONCLUSIONS HPLC-MS/MS provides a viable alternative to flow-injection MS/MS for the quantification of lysosomal enzyme activities. It is possible to assay 9 lysosomal enzymes using 1 or 2 reaction buffers, thus minimizing the number of separate incubations necessary.

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Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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High-Throughput Assay of 9 Lysosomal Enzymes for Newborn Screening

Spáčil¹,

Tatipaka²,

Barcenas³

et al. 2013

Clinical Chemistry

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“…Downscaling the assay volume, for instance, by employing digital microfluidics (DMF) as recently described (Sista et al 2011), might reduce the costs, although the cost structure using disposable cartridges is not yet clear.…”

Section: Resultsmentioning

confidence: 99%

Newborn Screening for Hunter Disease: A Small-Scale Feasibility Study

et al. 2013

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“…Assays for lysosomal enzyme activity, including I2S activity, in dried blood spots that may be suitable for newborn screening have been developed (Gelb et al 2006;Wolfe et al 2011;Sista et al 2011). A pilot population-based newborn screening using any of these methods has not yet been reported.…”

Section: Discussionmentioning

confidence: 99%

Detection by Urinary GAG Testing of Mucopolysaccharidosis Type II in an At-Risk Spanish Population

et al. 2012

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Hunter syndrome (Mucopolysaccharidosis type II) is an inherited lysosomal storage disorder with potentially severe degenerative consequences. Clinical diagnosis is not easy, although biochemical confirmation is straightforward, and sometimes patients are diagnosed at a late age. It is widely believed, for inborn errors of metabolism in general, that early diagnosis and management is of paramount importance for improving the prognosis of the disease. The objective of this study was to identify specific populations at risk of suffering from Hunter syndrome. Urine samples were obtained from children between the ages of 0 to 18, belonging to known risk groups of mucopolysaccharidosis (MPS) type II, for the semiquantitative (GAG test) and quantitative determination of glycosaminoglycans (GAG). One case of Hunter syndrome was found among the 130 samples that were collected and analysed. This study supports the feasibility of early diagnosis and the usefulness of screening tests for MPS II in specific paediatric populations.

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Rapid, single-step assay for Hunter syndrome in dried blood spots using digital microfluidics

Cited by 26 publications

References 9 publications

High-Throughput Assay of 9 Lysosomal Enzymes for Newborn Screening

High-Throughput Assay of 9 Lysosomal Enzymes for Newborn Screening

Newborn Screening for Hunter Disease: A Small-Scale Feasibility Study

Detection by Urinary GAG Testing of Mucopolysaccharidosis Type II in an At-Risk Spanish Population

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