Abstract
Background: We performed counselling for prenatal diagnosis (PD) of haemoglobinopathies in 372 couples. Thirty-four out of 372 (9.1%) did not undergo PD: six due to spontaneous abortion; nine because it was too difficult to make a decision if PD was positive; 18 because counselling excluded the carrier status of one or both parents; and one because parental mutations were mild.
Methods: Eleven out of 338 (3.3%) couples underwent PD because they had a thalassaemic child; 106 (31.4%) were found to be at high risk during pre-conceptional screening; 221 (65.4%) because of familiarity. Of 523 PDs in 486 (92.9%), including six dichorionic twin pregnancies, PD was performed on DNA from chorionic villi (CV), and in 37 from amniocytes (7.1%). In 1/523 cases, PD was not completed because DNA from CV was not sufficient; in two cases single tandem repeat analysis revealed maternal contamination of foetal DNA; in 7/522 (1.3%) cases PD revealed non-paternity. In 435/522 (83.3%) cases, PD was performed using reverse dot-blot and ARMS; 34/522 (6.5%) required sequencing. In 53/522 (10.2%) cases it was necessary to test globin loci for large rearrangements.
Results: One hundred and twenty out of 522 (23.0%) PDs revealed an affected foetus. In all but two cases the couple interrupted pregnancy. In the six twin pregnancies PD revealed a normal and a carrier foetus (two cases), carrier status in both foetuses (two cases) and a carrier and an affected foetus (two cases). In these latter cases the couple planned selective interruption.
Conclusions: Our PD procedure is successful and reliable, and is useful in high-risk areas characterised by molecular heterogeneity.
Grants from Regione Campania (DGRC 1901/09) and from University of Naples Federico II are gratefully acknowledged. We thank Jean Ann Gilder (Scientific Communication srl) for editing the text.
Conflict of interest statement
Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
References
1. Weatherall DJ, Clegg JB, editors. The thalassaemia syndrome, 4th ed. Oxford: Blackwell Science, 2010.Search in Google Scholar
2. Rosatelli MC, Tuveri T, Scalas MT, Leoni GB, Sardu R, Faà V, et al. Molecular screening and fetal diagnosis of beta-thalassemia in the Italian population. Hum Genet 1992;89:585–9.10.1007/BF00221942Search in Google Scholar PubMed
3. Pagano L, Lacerra G, Camardella L, De Angioletti M, Fioretti G, Maglione G, et al. Haemoglobin Neapolis, beta 126 (H4) Val → Gly: a novel beta-chain variant associated with a mild beta-thalassemia phenotype and displaying anomalous stability features. Blood 1991;78:3070–5.10.1182/blood.V78.11.3070.3070Search in Google Scholar
4. Pagano L, Carbone V, Fioretti G, Viola A, Buffardi S, Rametta V, et al. Compound heterozygosity for Hb Lepore-Boston and Hb Neapolis (Dhonburi) (beta 126 (H4) Val→Gly) in a patient from Naples, Italy. Haemoglobin 1997;21:1–15.10.3109/03630269708997506Search in Google Scholar
5. Wong WS, Chan AY, Yip SF, Ma ES. Thalassemia intermedia due to co-inheritance of beta0/beta(+)-thalassemia and (--SEA) alpha-thalassemia/Hb Westmead [alpha122 (H5) His→Gln (alpha2)] in a Chinese family. Haemoglobin 2004;28:151–6.10.1081/HEM-120035917Search in Google Scholar
6. Grosso M, Rescigno G, Zevino C, Matarazzo M, Poggi V, Izzo P. A rare case of compound heterozygosity for d+27 and Hb Neapolis (Dhonburi) associated to an atypical beta-thalassemia phenotype. Haematologica 2001;86:985–6.Search in Google Scholar
7. Cao A, Moi P. Genetic modifying factors in beta-thalassemia. Clin Chem Lab Med 2000;38:123–32.10.1515/CCLM.2000.019Search in Google Scholar PubMed
8. Weatherall DJ. Phenotype-genotype relationships in monogenic disease: lessons from the thalassaemias. Nature Rev 2001;2:245–55.10.1038/35066048Search in Google Scholar
9. Thein SL. Genetic modifiers of beta-thalassemia. Hum Mol Genet 2005;90:649–60.Search in Google Scholar
10. Maruotti GM, Frisso G, Calcagno G, Fortunato G, Castaldo G, Martinelli P, et al. Prenatal diagnosis of inherited diseases: the 20 years experience of an Italian Regional Reference Centre. Clin Chem Lab Med 2013;51:2211–7.10.1515/cclm-2013-0194Search in Google Scholar
11. Esposito G, Grosso M, Gottardi E, Izzo P, Camaschella C, Salvatore F. A unique origin for the Sicilian (deltabeta)°-thalassemia in 33 unrelated families and its rapid diagnostic characterization by PCR analysis. Hum Genet 1994;93:691–3.10.1007/BF00201572Search in Google Scholar
12. Camaschella C, Alfarano A, Gottardi E, Travi M, Primignani P, Caligaris Cappio F, et al. Prenatal diagnosis of fetal haemoglobin Lepore-Boston disease on maternal peripheral blood. Blood 1990;75:2102–6.10.1182/blood.V75.11.2102.2102Search in Google Scholar PubMed
13. Kattamis AC, Camaschella C, Sivera P, Surrey S, Fortina P. Human alpha-thalassemia syndromes: detection of molecular defects. Am J Haematol 1996;53:81–9110.1002/(SICI)1096-8652(199610)53:2<81::AID-AJH5>3.0.CO;2-#Search in Google Scholar
14. Galanello R, Origa R. Beta-thalassemia. Orph J Rare Dis 2010;5:11.10.1186/1750-1172-5-11Search in Google Scholar PubMed PubMed Central
15. Harteveld CL, Higgs DR. Alpha-thalassaemia. Orph J Rare Dis 2010;5:13.10.1186/1750-1172-5-13Search in Google Scholar PubMed PubMed Central
16. Higgs DR, Gibbons RJ. The molecular basis of alpha-thalassemia: a model for understanding human molecular genetics. Haematol Oncol Clin North Am 2010;24:1033–54.10.1016/j.hoc.2010.08.005http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000285267900004&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed
17. Chong SS, Boehm CD, Higgs DR, Cutting GR. Single-tube multiplex-PCR screen for common deletional determinants of alpha-thalassemia. Blood 2000;95:360–2.10.1182/blood.V95.1.360.001k03_360_362Search in Google Scholar PubMed
18. Tan AS, Quah TC, Low PS, Chong SS. A rapid and reliable 7-deletion multiplex polymerase chain reaction assay for alpha-thalassemia. Blood 2001;98:250–1.10.1182/blood.V98.1.250Search in Google Scholar PubMed
19. Moi P, Paglietti E, Sanna A, Brancati C, Tagarelli A, Galanello R, et al. Delineation of the molecular basis of delta- and normal HbA2 beta-thalassemia. Blood 1988;72:530–3.10.1182/blood.V72.2.530.530Search in Google Scholar
20. Colah RB, Surve R, Sawant P, Di Souza E, Italia K, Phanasgaonkar S, et al. HPLC studies in haemoglobinopathies. Ind J Pediatr 2007;74:657–62.10.1007/s12098-007-0117-8Search in Google Scholar PubMed
21. Rosatelli MC, Saba L. Prenatal diagnosis of beta-thalassemia and haemoglobinopathies. Med J Haematol Infect Dis 2009. Available from: http://www.mjhid.org/article/view/5079. Accessed on November 15, 2009.10.4084/MJHID.2009.011Search in Google Scholar PubMed PubMed Central
22. Grosso M, Palumbo I, Morelli E, Puzone S, Sessa R, Izzo P. Defective mRNA levels are responsible for a beta-thalassemia phenotype associated with Hb Federico II, a novel haemoglobin variant [beta 106 (G8) Leu∏Val]. Haematologica 2008;93:1096–8.10.3324/haematol.11722http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000257421200024&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed
23. Sessa R, Puzone S, Ammirabile M, Piscopo C, Pagano L, Colucci S, et al. Identification and molecular characterization of the Campania deletion, a novel alpha (0)-thalassemic defect, in two unrelated Italian families. Am J Haematol 2010;85:143–4.Search in Google Scholar
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