THE USE OF HPLC AS A TOOL FOR NEONATAL CORD BLOOD SCREENING OF HAEMOGLOBINOPATHY - A VALIDATION STUDY

Main Article Content

A. Al-Madhani
Anil Pathare
Salam Alkindi
(*) Corresponding Author:

Abstract

Background: Newborn cord blood screening identifies infants with underlying haemoglobinopathies before they develop the characteristic symptoms or sequelae. 

Aims: This study was performed to validate the interpretation high-performance chromatography (HPLC) along with complete blood count (CBC) results as a tool for universal neonatal screening of hemoglobin disorders in Oman. 

Methods: HPLC and CBC data on subjects who participated in the National Neonatal screening program at birth were obtained from archival records. The results recorded at birth were compared with a second study performed on the same subjects, after approval from the local medical research and ethics committee.

Results: Only 290 subjects from amongst the original cohort of 3740 newborns could be recalled between April 2010 to March 2011, to repeat HPLC and CBC, as well as perform confirmatory DNA studies, wherever necessary. All these subjects had been documented to show an initial abnormal result. 31 cases who had no HbA at birth on HPLC were confirmed as either homozygous β-thalassaemia major (n=5 subjects) or homozygous sickle cell anemia (n=26 subjects) by appropriate DNA analysis. Additionally, amongst 151 subjects, 72 subjects were studied in the initial study by Hb Bart’s quantitation using aalpha thalassaemia short program at birth. In this cohort, 42 subjects with Hb Bart’s >1% at birth could be confirmed as having either deletional or non-deletional thalassaemia by GAP PCR studies. No case of HbH was detected in this cohort. Further, carrier status for structural hemoglobin variants (HbS, HbC, HbD, HbE) (n=67) and beta thalassaemia allele with low HbA at birth (n=29 out of 41) were confirmed by relevant molecular studies.

Conclusions: The study validated the earlier observation by 100% concordance with results of CBC and HPLC. Presence of Hb Bart’s at birth does not always mean the presence of alpha thalassemia, as subjects with Hb Bart’s was below 1% by quantitation, were shown to be normal by molecular studies.

 

Key Words: Neonatal, screening, HPLC validation, haemoglobinopathy, sickle cell disease, thalassaemia

 


Downloads month by month

Downloads

Download data is not yet available.

Article Details

References

1. Leikin SL, Gallagher D, Kinney TR, Sloane D, Klug P, Rida W. Mortality in children and adolescents with sickle cell disease. Cooperative Study of Sickle Cell Disease. Pediatrics. 1989; 84:500-8. PMID: 2671914.
2. Serjeant GR. Natural history and determinants of clinical severity of sickle cell disease. Curr Opin Hematol. 1995; 2:103-8. PMID: 9371979
3. Koko J, Dufillot D, M’Ba-Meyo J, Gahouma D, Kani F. Mortality of children with sickle cell disease in a pediatric department in Central Africa. Arch Pediatr. 1998; 5:965-9. PMID: 9789626
4. Thomas C, Lemerle S, Bernaudin F, Feingold J, Guillou-Bataille M, Reinert P. Sickle cell anemia: study of the pediatric mortality in Ile de France from 1985 to 1992. Arch Pediatr. 1996; 3:445-51. PMID: 8763714
5. Platt OS, Brambilla DJ, Rosse WF, Milner PF, Castro O, Steinberg MH, et al. Mortality in sickle cell disease. Life expectancy and risk factors for early death. N Engl J Med. 1994; 330:1639-44. DOI: 10.1056/NEJM199406093302303.
6. Quinn CT. Sickle cell disease in childhood: from newborn screening through transition to adult medical care. Pediatr Clin North Am. 2013 Dec;60(6):1363-81. doi: 10.1016/j.pcl.2013.09.006
7. Riddington C, Owusu-Ofori S. Prophylactic antibiotics for preventing pneumococcal infection in children with sickle cell disease. Cochrane Database Syst Rev.;2002, (3):CD003427. DOI: 10.1002/14651858.CD00342.
8. Buchanan GR, Smith SJ. Pneumococcal septicemia despite pneumococcal vaccine and prescription of penicillin prophylaxis in children with sickle cell anemia. Am J Dis Child,1986, 140:428–432. PMID: 3962935.
9. Vichinsky E, Hurst D, Earles A, Kleman K, Lubin B Newborn screening for sickle cell disease: effect on mortality. Pediatrics.;1988, 81(6):749-55. PMID: 3368274.
10. King L, Fraser R, Forbes M, Grindley M, Ali S, Reid M. Newborn sickle cell disease screening: the Jamaican (1995-2006). J Med Screen. 2007;14(3):117-22. DOI: 10.1258/096914107782066185.
11. Alkindi S, Al Zadjali S, Al Madhani A, Daar S, Al Haddabi H, Al Abri Q, et al. Forecasting hemoglobinopathy burden through neonatal screening in Omani neonates. Hemoglobin.2010 Jan;34(2):135-44. DOI: 10.3109/03630261003677213.
12. Alkindi SS, Alzadjali S, Daar S, Sindhuvi E, Wali Y, Pathare AV, et al. A stepwise α-thalassemia screening strategy in high-prevalence areas. experience (1995-2006). Eur J Haematol. 2013 Aug;91(2):164-9. DOI: 10.1111/ejh.12136.
13. Jassim N, Al-Arrayed S, Gerard N, Al-Mukharraq H, Al-Ajami A, Ramasawmy R, et al. A mismatched-primer polymerase chain reaction-restriction fragment length polymorphism strategy for rapid screening of the polyadenylation signal mutation alpha(T-Saudi) (AATAAA-->AATAAG) in the alpha2-globin gene. Hemoglobin. 1999 Aug;23(3):213-20.
14. Rajab A, Patton MA. A study of consanguinity in the Sultanate of Oman. Ann Hum Biol. 2000 May-Jun;27(3):321-6. PMID: 10834296.
15. Alkindi S, Pathare A, Al-Madhani A, Al-Zadjali S, Al-Haddabi H, Al-Abri Q, et al. Neonatal Screening: Mean haemoglobin and red cell indices in cord blood from Omani neonates. Sultan Qaboos Univ Med J.2011, 11(4):462-9. PMID: 22087394 PMCID: PMC3206748.
16. Piel FB, Hay SI, Gupta S, Weatherall DJ, Williams TN. Global burden of sickle cell anaemia in children under five, 2010-2050: modelling based on demographics, excess mortality, and interventions. PLoS Med. 2013;10(7): e1001484. DOI: 10.1371/journal.pmed.1001484.
17. Higgs DR, Vickers MA, Wilkie AO, Pretorius IM, Jarman AP, Weatherall DJ.: A review of the molecular genetics of the human a-globulin gene cluster. Blood 1989; 73(5):1091-1108. PMID: 2649166.
18. Weatheral DJ. The thalassaemias. In: Beutler E, Litchmen MA, Coller BS, Kipps TJ, editors. William’s Hematology. 5th Edition (International edition). New York: McGraw-Hill Inc, Health Professions Division, 1995:581-615.
19. Nasserullah Z, Al-Jame A, Abu Srair H, Al Qatari G, Al Naim S, Al Aqib A et al. Neonatal screening for sickle cell disease, glucose-6-phosphate dehydrogenase deficiency and -thalassaemia in Qatif and Al Hassa. Ann Saudi Med:1998; 18(4):289-92. PMID: 17344674.
20. White JM, Christie BS, Nam D, Daar S, Higgs DR. Frequency and clinical significance of erythrocyte genetic abnormalities in Omanis. J Med. Genet. 1993; 30(5):396-400. PMID: 8320702 PMCID: PMC1016376.