The Utilization of MS-MLPA as the First-Line Test for the Diagnosis of Prader–Willi Syndrome in Thai Patients

Abstract

Prader–Willi syndrome (PWS) is a genetic disorder caused by the expression disruption of genes on the paternally inherited allele of chromosome 15q11.2-q13. Apart from clinical diagnostic criteria, PWS is confirmed by genetic testing. Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) is one of the molecular techniques used to analyze this syndrome. This study aimed to evaluate the concordance of the test results of MS-MLPA with conventional techniques in the diagnosis of PWS in Thai patients. Forty leftover specimens from routine genetic testing (MS-PCR and FISH) were tested to obtain MS-MLPA results. By comparison, perfect concordance was shown between the result of MS-MLPA and those of conventional techniques. In conclusion, MS-MLPA is an accurate and cost-effective assay that can be used to confirm PWS diagnosis with explicit deletion of affected genes.

Keywords

PWS - MS-MLPA - Imprinting disorder

Authors' Contributions

C.P. designed the study, collected data, performed genetic testing, and wrote the manuscript; P.O., V.K., and B.R. supervised genetic testing and provided samples; D.W. was an attending physician, who supervised and provided samples and clinical data; and T.K. designed the study, supervised for data analysis, and revised the manuscript.

Publication History

Received: 04 August 2021

Accepted: 22 November 2021

Article published online:
07 January 2022

• References

• 1 Cassidy SB, Schwartz S, Miller JL, Driscoll DJ. Prader-Willi syndrome. Genet Med 2012; 14 (01) 10-26
• 2 Angulo MA, Butler MG, Cataletto ME. Prader-Willi syndrome: a review of clinical, genetic, and endocrine findings. J Endocrinol Invest 2015; 38 (12) 1249-1263
• 3 Miller JL, Lynn CH, Driscoll DC. et al. Nutritional phases in Prader-Willi syndrome. Am J Med Genet A 2011; 155A [phone]
• 4 Lewis BA, Freebairn L, Heeger S, Cassidy SB. Speech, and language skills of individuals with Prader-Willi syndrome. Am J Speech Lang Pathol 2002; 11: 285-294
• 5 Whittington J, Holland A, Webb T, Butler J, Clarke D, Boer H. Academic underachievement by people with Prader-Willi syndrome. J Intellect Disabil Res 2004; 48 (Pt 2): 188-200
• 6 Harris RM, Stafford DEJ. Prader Willi syndrome: endocrine updates and new medical therapies. Curr Opin Endocrinol Diabetes Obes 2020; 27 (01) 56-62
• 7 Proffitt J, Osann K, McManus B. et al. Contributing factors of mortality in Prader-Willi syndrome. Am J Med Genet A 2019; 179 (02) 196-205
• 8 Glenn CC, Driscoll DJ, Yang TP, Nicholls RD. Genomic imprinting: potential function and mechanisms revealed by the Prader-Willi and Angelman syndromes. Mol Hum Reprod 1997; 3 (04) 321-332
• 9 Bittel DC, Butler MG. Prader-Willi syndrome: clinical genetics, cytogenetics and molecular biology. Expert Rev Mol Med 2005; 7 (14) 1-20
• 10 Cassidy SB, Driscoll DJ. Prader-Willi syndrome. Eur J Hum Genet 2009; 17 (01) 3-13
• 11 Butler MG, Miller JL, Forster JL. Prader-Willi Syndrome - clinical genetics, diagnosis and treatment approaches: an update. Curr Pediatr Rev 2019; 15 (04) 207-244
• 12 MRC Holland Product description ME028–C1 Prader-Willi/Angelman-vC1–04. Available at: https://www.mrcholland.com/product/ME028/397
• 13 MS-MLPA-General-Protocol-MSP-v011. Available at: https://support.mrcholland.com/downloads/files/ms-mlpa-general-protocol-one-tube
• 14 Bittel DC, Kibiryeva N, Butler MG. Methylation-specific multiplex ligation-dependent probe amplification analysis of subjects with chromosome 15 abnormalities. Genet Test 2007; 11 (04) 467-475
• 15 Dikow N, Nygren AO, Schouten JP. et al. Quantification of the methylation status of the PWS/AS imprinted region: comparison of two approaches based on bisulfite sequencing and methylation-sensitive MLPA. Mol Cell Probes 2007; 21 (03) 208-215
• 16 Henkhaus RS, Kim SJ, Kimonis VE. et al. Methylation-specific multiplex ligation-dependent probe amplification and identification of deletion genetic subtypes in Prader-Willi syndrome. Genet Test Mol Biomarkers 2012; 16 (03) 178-186
• 17 Botezatu A, Puiu M, Cucu N. et al. Comparative molecular approaches in Prader-Willi syndrome diagnosis. Gene 2016; 575 (2 Pt 1): 353-358
• 18 Brown A, Flintoff K, Felix C. et al. Prader-Willi/Angelman syndrome: a comparative study of MS-PCR and MS-MLPA. Med Res Arch 2017;5(9). Accessed November 20, 2020 at https://esmed.org/MRA/mra/article/view/1437
• 19 Mahmoud R, Singh P, Weiss L. et al. Newborn screening for Prader-Willi syndrome is feasible: early diagnosis for better outcomes. Am J Med Genet A 2019; 179 (01) 29-36
• 20 Wiriyaukaradecha S, Patmasiriwat P, Wasant P, Tantiniti P. Molecular markers for diagnosis of Prader-Willi syndrome in Thai patients by fish. Southeast Asian J Trop Med Public Health 2003; 34 (04) 881-886
• 21 Pangkanon S. Molecular diagnosis of Prader-Willi syndrome. J Med Assoc Thai 2003; 86 (Suppl. 03) S510-S516
• 22 Promkan M, Teingtat S, Stheinkijkarnchai A, Wasant P, Patmasiriwat P. Highest accuracy of combined consensus clinical criteria and SNRPN gene molecular markers in diagnosis of Prader-Willi syndrome in Thai patients. Clin Chem Lab Med 2007; 45 (08) 972-980
• 23 Driscoll DJ, Miller JL, Schwartz S. et al. Prader-Willi Syndrome. In: Adam MP, Ardinger HH, Pagon RA. et al. eds. GeneReviews® [Internet]. Seattle, WA: University of Washington; 1993. –2021. Accessed January 29, 2021 at: https://www.ncbi.nlm.nih.gov/books/NBK1330/
• 24 Butler MG, Bittel DC, Kibiryeva N, Talebizadeh Z, Thompson T. Behavioral differences among subjects with Prader-Willi syndrome and type I or type II deletion and maternal disomy. Pediatrics 2004; 113 (3 Pt 1): 565-573
• 25 Zarcone J, Napolitano D, Peterson C. et al. The relationship between compulsive behaviour and academic achievement across the three genetic subtypes of Prader-Willi syndrome. J Intellect Disabil Res 2007; 51 (Pt. 6): 478-487
• 26 Milner KM, Craig EE, Thompson RJ. et al. Prader-Willi syndrome: intellectual abilities and behavioural features by genetic subtype. J Child Psychol Psychiatry 2005; 46 (10) 1089-1096
• 27 Varela MC, Kok F, Setian N, Kim CA, Koiffmann CP. Impact of molecular mechanisms, including deletion size, on Prader-Willi syndrome phenotype: study of 75 patients. Clin Genet 2005; 67 (01) 47-52
• 28 Butler MG. Clinical and genetic aspects of the 15q11.2 BP1-BP2 microdeletion disorder. J Intellect Disabil Res 2017; 61 (06) 568-579
• 29 Rafi SK, Butler MG. The 15q11.2 BP1-BP2 microdeletion (Burnside-Butler) syndrome: in silico analyses of the four coding genes reveal functional associations with neurodevelopmental phenotypes. Int J Mol Sci 2020; 21 (09) 3296
• 30 Baldwin I, Shafer RL, Hossain WA. et al. Genomic, clinical, and behavioral characterization of 15q11.2 BP1-BP2 deletion (Burnside-Butler) syndrome in five families. Int J Mol Sci 2021; 22 (04) 1660
• 31 Beygo J, Buiting K, Ramsden SC, Ellis R, Clayton-Smith J, Kanber D. Update of the EMQN/ACGS best practice guidelines for molecular analysis of Prader-Willi and Angelman syndromes. Eur J Hum Genet 2019; 27 (09) 1326-1340
• 32 Goldstone AP, Holland AJ, Hauffa BP, Hokken-Koelega AC, Tauber M. speakers contributors at the Second Expert Meeting of the Comprehensive Care of Patients with PWS. Recommendations for the diagnosis and management of Prader-Willi syndrome. J Clin Endocrinol Metab 2008; 93 (11) 4183-4197
• 33 McCandless SE. Committee on Genetics. Clinical report—health supervision for children with Prader-Willi syndrome. Pediatrics 2011; 127 (01) 195-204
• 34 Deal CL, Tony M, Höybye C, Allen DB, Tauber M, Christiansen JS. 2011 Growth Hormone in Prader-Willi Syndrome Clinical Care Guidelines Workshop Participants. GrowthHormone Research Society workshop summary: consensus guidelines for recombinant human growth hormone therapy in Prader-Willi syndrome. J Clin Endocrinol Metab 2013; 98 (06) E1072-E1087
• 35 Muscogiuri G, Formoso G, Pugliese G, Ruggeri RM, Scarano E, Colao A. RESTARE. Prader-Willi syndrome: An up to date on endocrine and metabolic complications. Rev Endocr Metab Disord 2019; 20 (02) 239-250