Reliability of enzyme assays in dried blood spots for diagnosis of 4 lysosomal storage disorders

Ceci, Romina; Francesco, Pablo N. De; Mucci, Juan Marcos; Cancelarich, Lorena N.; Fossati, Carlos A.; Rozenfeld, Paula

doi:10.4236/abc.2011.13008

Cited by 16 publications

(23 citation statements)

References 21 publications

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“…The intra and inter-assay CVs ranges were 6.9-8.9% and 7.6-12%, respectively. These results are similar to those obtained by Ceci et al using a GBA enzymatic assay in microplates [12].…”

Section: Precision and Recoverysupporting

confidence: 92%

“…However, early diagnosis of GD may enable the early treatment of patients before the onset of irreversible damage, which will be particularly important for those patients who will go on developing neuropathic disease excepted GD type 2 [26][27][28]. A practical way to achieve an early diagnosis is through a newborn screening program, as has been suggested by other authors [12,19]. Selection of an appropriate enzyme assay in order to detect infants at risk of having GD, is of great importance in making a clear diagnosis.…”

Section: Discussionmentioning

confidence: 99%

“…Chamoles et al reported in 30 newborn DBS samples a range of enzymatic activity 2.65-11.14 mol/l blood/h with a mean value 5.27 mol/l blood/h [19] while Civallero et al obtained an enzymatic activity range 1.92-8.77 mol/l blood/h with a mean value of 4.93 mol/l blood/h in 25 newborn DBS samples [11]. Ceci et al reported a cut off value of 1.8 mol/l blood/h [12]. Through the use of this fluorometric assay, the definition of reference intervals in GD patients, carriers and in healthy individuals in our population will be possible.…”

Section: Discussionmentioning

confidence: 99%

“…The diagnosis of GD could be done by using several analytical techniques: enzymatic assay [10][11][12], mass spectrometry [13][14][15], immune-quantification [16] or molecular biology [17,18]. Chamoles et al described for the first time assays for the determination of lysosomal enzymes activities in dried blood spots on filter paper (DBS) from newborns screening cards [19].…”

Section: Introductionmentioning

confidence: 99%

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Ultramicro-fluorometric assay for the diagnosis of Gaucher disease in dried blood spots on filter paper

Herrera

Monaga

Campos

et al. 2013

Journal of Neonatal-Perinatal Medicine

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BACKGROUND: Gaucher disease (GD) is a lysosomal storage disorder characterized by a deficiency of the lysosomal acid ␤-D-glucosidase (GBA). The aim of this study was to develop an ultramicro-fluorometric assay based on the method of Chamoles et al. for determining GBA activity in dried blood spots on filter paper (DBS). METHODS: The assay used 3-mm diameter blood spot and 8 mmol/l of 4-methylumbelliferyl-␤-D-glucoside as enzymatic substrate. The reaction occurred in plates incubated at 37 • C for 20 hours and the enzyme activity was expressed in mol hydrolysed substrate/l blood/h. The fluorescence of the enzyme product was automatically measured in a fluorometer-photometer reader (SUMA Technology). RESULTS: The intra and inter-assay coefficients of variation were lower than 9 and 12%, respectively, and the recovery range was 97-109%.Three patients with GD were correctly diagnosed using the ultramicroassay. Healthy newborn DBS samples (n = 3003) from the National Neonatal Screening Program were analyzed, and the mean GBA activity was 5.7 mol/l blood/h. Our assay showed high Pearson (n = 26; r = 0.99) and concordance correlations (ρc = 0.99) with the traditional method described by Chamoles et al. CONCLUSIONS:The analytical performance characteristics of our ultramicro-fluorometric assay suggest that it can be used in the diagnosis of GD in newborns and adults.

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“…The intra and inter-assay CVs ranges were 6.9-8.9% and 7.6-12%, respectively. These results are similar to those obtained by Ceci et al using a GBA enzymatic assay in microplates [12].…”

Section: Precision and Recoverysupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Ultramicro-fluorometric assay for the diagnosis of Gaucher disease in dried blood spots on filter paper

Herrera

Monaga

Campos

et al. 2013

Journal of Neonatal-Perinatal Medicine

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“…This finding is not surprising, as LAL is an enzyme potentially expressed by any cellular lines containing lysosomes—both white blood cells [18] and platelets [19] possess lysosomal structures.…”

Section: Discussionmentioning

confidence: 99%

A Relative Deficiency of Lysosomal Acid Lypase Activity Characterizes Non-Alcoholic Fatty Liver Disease

Tovoli

Napoli

Negrini

et al. 2017

IJMS

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Lysosomal acid lipase (LAL) is a key enzyme in lipid metabolism. Initial reports have suggested a role for a relative acquired LAL deficiency in non-alcoholic fatty liver disease (NAFLD)—however, it is still unclear whether this mechanism is specific for NAFLD. We aimed to determine LAL activity in a cohort of NAFLD subjects and in a control group of hepatitis C virus (HCV)-infected patients, investigating the role of liver cirrhosis. A total of 81 patients with a diagnosis of NAFLD, and 78 matched controls with HCV-related liver disease were enrolled. For each patient, LAL activity was determined on peripheral dried blood spots (DBS) and correlated with clinical and laboratory data. A subgroup analysis among cirrhotic patients was also performed. LAL activity is significantly reduced in NAFLD, compared to that in HCV patients. This finding is particularly evident in the pre-cirrhotic stage of disease. LAL activity is also correlated with platelet and white blood cell count, suggesting an analytic interference of portal-hypertension-induced pancytopenia on DBS-determined LAL activity. NAFLD is characterized by a specific deficit in LAL activity, suggesting a pathogenetic role of LAL. We propose that future studies on this topic should rely on tissue specific analyses, as peripheral blood tests are also influenced by confounding factors.

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Twenty years of Colombian experience with enzymatic screening in patients with features of mucopolysaccharidosis

2022

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Mucopolysaccharidoses (MPSs) are a group of genetic alterations whose effect is the progressive intralysosomal accumulation of glycosaminoglycans. Affected individuals are deficient in one or more lysosomal enzymes which, depending on the MPS, may cause coarse facial features, short stature, multiple skeletal dysplasia, joint stiffness, or developmental delay. Their diagnosis is mostly performed late or incorrectly, and it represents a challenge since it requires specialized tests only performed in major cities. This makes it difficult for patients to have access to physicians since their geographical location is distant and therefore, the use of samples collected in solid‐phase represents an advantage for the study of high‐risk populations. In addition, epidemiological information about rare diseases, especially in Latin America, is scarce or inconsistent. Our aim was to report the experience of 20 years of selective screening by assessing enzyme activity and reporting incidence values of MPS in Colombia. This study validated a group of fluorometric endpoint techniques in 8239 patients. The samples were dried blood spots (DBS) collected on filter paper and leukocyte extracts. Reference values in the Colombian population for α‐ l ‐iduronidase, iduronate 2‐sulfatase, α‐ N ‐acetylglucosaminidase, N ‐acetylglucosamine‐6‐sulfate sulfatase, β‐galactosidase, arylsulfatase B, and β‐glucuronidase were established in leukocyte extracts, and patients reference ranges were updated in the case of DBS samples. Incidence values were calculated for each MPS and the distribution of cases across the country is also shown. This study offers very useful information for the health system, the scientific community, and it facilitates the diagnosis of these disorders. This is indispensable when seeking to develop new diagnostic or treatment approaches for patients.

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Reliability of enzyme assays in dried blood spots for diagnosis of 4 lysosomal storage disorders

Cited by 16 publications

References 21 publications

Ultramicro-fluorometric assay for the diagnosis of Gaucher disease in dried blood spots on filter paper

Ultramicro-fluorometric assay for the diagnosis of Gaucher disease in dried blood spots on filter paper

A Relative Deficiency of Lysosomal Acid Lypase Activity Characterizes Non-Alcoholic Fatty Liver Disease

Twenty years of Colombian experience with enzymatic screening in patients with features of mucopolysaccharidosis

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