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Development of a fluorometric microtiter plate-based enzyme assay for arylsulfatase B (MPS VI) using dried blood spots

View Article: PubMed Central - PubMed

ABSTRACT

Mucopolysaccharidosis type VI or Maroteaux–Lamy syndrome is an autosomal recessive lysosomal storage disorder caused by deficiency of arylsulfatase B (ARS-B) enzyme activity. It results in mild to severe multi-organ system failure from accumulation of undigested glycosaminoglycans (GAGs); dermatan sulfate and chondroitin-4-sulfate. We have developed a single-step enzyme assay using a fluorescent substrate and dried blood spots to measure ARS-B activity to identify disease patients. This assay is robust, reproducible, specific and convenient to perform.

No MeSH data available.


Related in: MedlinePlus

A) ARS-B enzyme activity comparison between normal control (circles) and MPS VI affected (squares) dried blood spots. Error bars show mean and SEM. B) Stability of ARS-B activity in dried-blood spots stored at different temperatures and durations (initial activity; − 20 °C, 4 °C & 22 °C for 7 days; 30 °C for 6 h; 37 °C for 2 h).
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f0005: A) ARS-B enzyme activity comparison between normal control (circles) and MPS VI affected (squares) dried blood spots. Error bars show mean and SEM. B) Stability of ARS-B activity in dried-blood spots stored at different temperatures and durations (initial activity; − 20 °C, 4 °C & 22 °C for 7 days; 30 °C for 6 h; 37 °C for 2 h).

Mentions: Fig. 1A shows the relative ARS-B enzyme activity in the DBS tested from normal and known MPS VI patients. The Duke-BGL control DBS (gray circles) showed an enzyme activity range of 21.6–103.6 pmol/punch*h. The calculated mean ± standard deviation values were 49.3 ± 15.6 pmol/punch*h. The MPS VI-affected DBS (black squares) showed enzyme activity ranging between 0.7–8.9 pmol/punch*h with a mean value of 4.8 ± 2.8 (mean ± std. dev.). The ARS-B deficient DBS activity levels were clearly separated and distinguishable from those of normal control spots (p < 0.0001, Tukey's t-test).


Development of a fluorometric microtiter plate-based enzyme assay for arylsulfatase B (MPS VI) using dried blood spots
A) ARS-B enzyme activity comparison between normal control (circles) and MPS VI affected (squares) dried blood spots. Error bars show mean and SEM. B) Stability of ARS-B activity in dried-blood spots stored at different temperatures and durations (initial activity; − 20 °C, 4 °C & 22 °C for 7 days; 30 °C for 6 h; 37 °C for 2 h).
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5121359&req=5

f0005: A) ARS-B enzyme activity comparison between normal control (circles) and MPS VI affected (squares) dried blood spots. Error bars show mean and SEM. B) Stability of ARS-B activity in dried-blood spots stored at different temperatures and durations (initial activity; − 20 °C, 4 °C & 22 °C for 7 days; 30 °C for 6 h; 37 °C for 2 h).
Mentions: Fig. 1A shows the relative ARS-B enzyme activity in the DBS tested from normal and known MPS VI patients. The Duke-BGL control DBS (gray circles) showed an enzyme activity range of 21.6–103.6 pmol/punch*h. The calculated mean ± standard deviation values were 49.3 ± 15.6 pmol/punch*h. The MPS VI-affected DBS (black squares) showed enzyme activity ranging between 0.7–8.9 pmol/punch*h with a mean value of 4.8 ± 2.8 (mean ± std. dev.). The ARS-B deficient DBS activity levels were clearly separated and distinguishable from those of normal control spots (p < 0.0001, Tukey's t-test).

View Article: PubMed Central - PubMed

ABSTRACT

Mucopolysaccharidosis type VI or Maroteaux&ndash;Lamy syndrome is an autosomal recessive lysosomal storage disorder caused by deficiency of arylsulfatase B (ARS-B) enzyme activity. It results in mild to severe multi-organ system failure from accumulation of undigested glycosaminoglycans (GAGs); dermatan sulfate and chondroitin-4-sulfate. We have developed a single-step enzyme assay using a fluorescent substrate and dried blood spots to measure ARS-B activity to identify disease patients. This assay is robust, reproducible, specific and convenient to perform.

No MeSH data available.


Related in: MedlinePlus