Objectives: The need for early detection of lysosomal storage diseases (LSDs) for which therapeutic options are available makes them attractive candidate disorders to perform high-throughput population screening. This is a pilot study designed to simultaneously screen for Krabbe, Niemann-Pick types A/B, Fabry, Gaucher and Pompe diseases in putatively normal Indian subjects, using dried blood spots and a liquid chromatography-tandem mass spectrometry method. Methods: Blood spots from 12,559 putatively normal subjects were used to measure 5 lysosomal enzymes: galactocerebrosidase, acid sphingomyelinase, α-galactosidase, β-glucocerebrosidase, and α-glucosidase. From each blood spot, 3.2-mm punches were extracted and incubated using specific substrates and internal standards. After several liquid- and solid-phase extraction steps, the resulting solution was reconstituted and injected into a triple quadrupole, liquid chromatography-tandem mass spectrometer after recombining the reaction products into a single 96-well plate. Results: A standard calibration curve demonstrated good linearity for each enzyme. No positive case was detected among the 12,559 putatively normal subjects tested. Conclusion: Tandem mass spectrometry technology makes it possible to perform high-throughput screening to identify LSDs using blood spots. Further large-scale studies to determine the population prevalence and incidence of these disorders are warranted.
1. Fletcher JM. Screening for lysosomal storage disorders--a clinical perspective. J Inherit Metab Dis 2006;29(2-3):405–8.
2. Vitner EB, Platt FM, Futerman AH. Common and uncommon pathogenic cascades in lysosomal storage diseases. J Biol Chem 2010;285(27):20423–7.
3. Parkinson-Lawrence EJ, Shandala T, Prodoehl M, Plew R, Borlace GN, Brooks DA. Lysosomal storage disease: revealing lysosomal function and physiology. Physiology (Bethesda) 2010;25(2):102–15.
4. Futerman AH, van Meer G. The cell biology of lysosomal storage disorders. Nat Rev Mol Cell Biol 2004;5(7):554–65.
5. Desnick RJ. Enzyme replacement and beyond. J Inherit Metab Dis 2001;24(2):251–65.
6. Prasad VK, Kurtzberg J. Transplant outcomes in mucopolysaccharidoses. Semin Hematol 2010;47(1):59–69.
7. Abdi M, Hakhamaneshi MS, Alaei MR, Azadi NA, Vakili R, Zamanfar D, et al. Determination of Biological Variance and Validation of a Fluorometric Assay for Measurement of α-l-Iduronidase Activity in Dried Blood Spots Samples: The First Experience in Iran. Indian J Clin Biochem 2015;30(3):318–22.
8. Li Y, Scott CR, Chamoles NA, Ghavami A, Pinto BM, Turecek F, et al. Direct multiplex assay of lysosomal enzymes in dried blood spots for newborn screening. Clin Chem 2004;50(10):1785–96.
9. Gelb MH, Turecek F, Scott CR, Chamoles NA. Direct multiplex assay of enzymes in dried blood spots by tandem mass spectrometry for the newborn screening of lysosomal storage disorders. J Inherit Metab Dis 2006;29(2-3):397–404.
10. Zhang XK, Elbin CS, Chuang WL, Cooper SK, Marashio CA, Beauregard C, et al. Multiplex enzyme assay screening of dried blood spots for lysosomal storage disorders by using tandem mass spectrometry. Clin Chem 2008;54(10):1725–8.
11. Spáčil Z, Elliott S, Reeber SL, Gelb MH, Scott CR, Tureček F. Comparative triplex tandem mass spectrometry assays of lysosomal enzyme activities in dried blood spots using fast liquid chromatography: application to newborn screening of Pompe, Fabry, and Hurler diseases. Anal Chem 2011;83(12):4822–8.
12. Orsini JJ, Morrissey MA, Slavin LN, Wojcik M, Biski C, Martin M, Keutzer J, Zhang XK, Chuang WL, Elbin C, Caggana M. Implementation of newborn screening for Krabbe disease: population study and cutoff determination. Clin Biochem 2009;42(9):877–84.
13. Marsden D, Levy H. Newborn screening of lysosomal storage disorders. Clin Chem 2010;56(7):1071–9.
14. Liao HC, Chiang CC, Niu DM, Wang CH, Kao SM, Tsai FJ, et al. Detecting multiple lysosomal storage diseases by tandem mass spectrometry--a national newborn screening program in Taiwan. Clin Chim Acta 2014;431:80–6.
15. Gelb MH, Scott CR, Turecek F. Newborn screening for lysosomal storage diseases. Clin Chem 2015;61(2):335–46.
16. Matern D, Gavrilov D, Oglesbee D, Raymond K, Rinaldo P, Tortorelli S. Newborn screening for lysosomal storage disorders. Semin Perinatol 2015;39(3):206–16.
17. Elliott S, Buroker N, Cournoyer JJ, Potier AM, Trometer JD, Elbin C, et al. Dataset and standard operating procedure for newborn screening of six lysosomal storage diseases: By tandem mass spectrometry. Data Brief 2016;8:915–24.
18. Jack RM, Gordon C, Scott CR, Kishnani PS, Bali D. The use of acarbose inhibition in the measurement of acid alpha-glucosidase activity in blood lymphocytes for the diagnosis of Pompe disease. Genet Med 2006;8(5):307–12.
19. Supriya M, De T, Christopher R. Age and gender-specific reference intervals for lysosomal enzymes in dried blood spot samples: A study in Indian population. Clin Biochem 2017;50(15):858–63.
20. Evaluation of precision performance of quantitative measurement methods; approved guideline—second edition. Available at: https://yeec.com/uploadimages1/forum/2013-2/201321114424796191.pdf. Accessed Apr 15, 2020.
21. Wang RY, Bodamer OA, Watson MS, Wilcox WR; ACMG Work Group on Diagnostic Confirmation of Lysosomal Storage Diseases. Lysosomal storage diseases: diagnostic confirmation and management of presymptomatic individuals. Genet Med 2011;13(5):457–84.
22. Verma IC. Burden of genetic disorders in India. Indian J Pediatr. 2000;67(12):893–8.
23. Kadali S, Kolusu A, Gummadi MR, Undamatla J. The relative frequency of lysosomal storage disorders: a medical genetics referral laboratory's experience from India. J Child Neurol 2014;29(10):1377–82.
24. Sheth J, Mistri M, Sheth F, Shah R, Bavdekar A, Godbole K, et al. Burden of lysosomal storage disorders in India: experience of 387 affected children from a single diagnostic facility. JIMD Rep 2014;12:51–63.
25. Verma PK, Ranganath P, Dalal AB, Phadke SR. Spectrum of Lysosomal storage disorders at a medical genetics center in northern India. Indian Pediatr 2012;49(10):799–804.
26. Agarwal S, Lahiri K, Muranjan M, Solanki N. The face of lysosomal storage disorders in India: a need for early diagnosis. Indian J Pediatr 2015;82(6):525–9.
27. Mechtler TP, Stary S, Metz TF, De Jesús VR, Greber-Platzer S, Pollak A, et al. Neonatal screening for lysosomal storage disorders: feasibility and incidence from a nationwide study in Austria. Lancet 2012;379(9813):335–41.
28. Wittmann J, Karg E, Turi S, Legnini E, Wittmann G, Giese AK,et al. Newborn screening for lysosomal storage disorders in hungary. JIMD Rep 2012;6:117–25.
29. Burlina AB, Polo G, Salviati L, Duro G, Zizzo C, Dardis A, et al. Newborn screening for lysosomal storage disorders by tandem mass spectrometry in North East Italy. J Inherit Metab Dis 2018;41(2):209–19.
30. Cho SE, Kwak JR, Lee H, Seo DH, Song J. Triplex tandem mass spectrometry assays for the screening of 3 lysosomal storage disorders in a Korean population. Clin Chim Acta 2016;454:20–7.
31. Elliott S, Buroker N, Cournoyer JJ, Potier AM, Trometer JD, Elbin C, et al. Pilot study of newborn screening for six lysosomal storage diseases using Tandem Mass Spectrometry. Mol Genet Metab 2016;118(4):304–9.
32. Burton BK, Charrow J, Hoganson GE, Waggoner D, Tinkle B, Braddock SR, et al G. Newborn Screening for Lysosomal Storage Disorders in Illinois: The Initial 15-Month Experience. J Pediatr 2017;190:130–5.
33. Scott CR, Elliott S, Buroker N, Thomas LI, Keutzer J, Glass M, et al. Identification of infants at risk for developing Fabry, Pompe, or mucopolysaccharidosis-I from newborn blood spots by tandem mass spectrometry. J Pediatr 2013;163(2):498–503.