Rapid Mid-Trimester Prenatal Diagnosis of Beta-thalassaemia and other Haemoglobinopathies using a Non-Radioactive Anion Exchange HPLC Technique—An Indian Experience

Rao, V. Divakara; Natrajan, P. G.; Lulla, Chander; Bandodkar, S. B.

doi:10.1002/(sici)1097-0223(199708)17:8<725::aid-pd134>3.0.co;2-8

Cited by 14 publications

(5 citation statements)

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“…Prenatal diagnosis of thalassemia at most centers is usually done using standard DNA analysis by polymerase chain reaction (PCR) of fetal tissues obtained by chrorionic villus sampling (CVS) between 8 and 12 wk of gestation, amniocentesis between 14 and 20 wk gestation or cordocentesis after 18 wk of gestation (2, 3). Analysis of fetal Hbs using automated HPLC system has been used as another alternative when DNA analysis is not routinely performed and mutations of the parents are unknown (4–6). Recently, it has been reported that a kit for analysis of Hb fractions and variants on the capillary electrophoresis system could be an alternative method for Hb analysis (7).…”

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confidence: 99%

Analysis of fetal blood using capillary electrophoresis system: a simple method for prenatal diagnosis of severe thalassemia diseases

Srivorakun

Fucharoen

Sae-ung

et al. 2009

European J of Haematology

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In Thailand where the total population is approximately 63 millions with 800 000 births per year, 30-40% of the peoples are carriers of thalassemia and hemoglobinopathies. These include 20-30% of a-thalassemia trait, 3-9% with b-thalassemia trait, 20-30% with Hb E (1). With such high prevalence, thalassemia syndromes are very common, leading to major health and socioeconomic problem of these countries. It is estimated that 1% of Thai population has thalassemia disease and each year there are more than 12 000 new births with thalassemia syndromes. The national prevention and control program of thalassemia has been implemented throughout the country. Three severe thalassemia diseases, targeted for prevention and control are Hb Bart's hydrops fetalis (homozygous a°-thalassemia), homozygous b-thalassemia and Hb E-b-thalassemia. If parents are both carriers, there is a 25% chance in each pregnancy that the child will be born with these thalassemia diseases. One objective of the prevention and control program is to prevent birth of new cases with these three severe thalassemia diseases. Carrier screening and prenatal diagnosis for couple at risk for these Abstract Introduction: Prenatal diagnosis of severe a-and b-thalasssemia diseases is usually performed by DNA analysis. Objective: To establish a simple method, we have evaluated the reliability of prenatal diagnosis by fetal blood analysis using automated capillary electrophoresis system. Methods: Forty-seven fetal blood specimens collected by cordocentesis at 18-28 wk of gestation were analyzed by the capillary electrophoresis system (Sebia). Fetal DNA was analyzed for respective thalassemia alleles by PCR. Results: Among 47 fetuses, 20 were at risk for the Hb Bart's hydrops fetalis. DNA analysis identified four cases of homozygous a°-thalassemia (SEA type). Hb analysis by the capillary electrophoresis demonstrated a major peak of Hb Bart's (78.4-81.3%), Hb H (0.8-1.4%) and minor peaks of presumably embryonic Hbs. No Hb F and Hb A was observed. The level of Hb Bart's was found to be 3.4-5.8% in unaffected heterozygote whereas normal fetus had no Hb Bart's. Among the remaining 27 fetuses at risk for Hb E-b-thalassemia, DNA analysis identified 12 affected fetuses. Hb analysis showed Hb F (94.9-98.9%) and Hb E (1.1-1.8%) without Hb A in all cases. The levels of Hb A were found to be (4.3-7.2%), (1.0-5.5%) and (2.1-3.9%) in normal, heterozygous Hb E and heterozygous b-thalassemia fetuses, respectively. Affected and unaffected fetuses could be easily distinguished. Conclusion: Capillary electrophoresis system is a simple and automated procedure for accurate prenatal diagnosis of severe thalassemia diseases which could readily be performed in routine setting.

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confidence: 99%

Analysis of fetal blood using capillary electrophoresis system: a simple method for prenatal diagnosis of severe thalassemia diseases

Srivorakun

Fucharoen

Sae-ung

et al. 2009

European J of Haematology

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“…However, in most laboratories, HPLC has been used for diagnosis of Hb variants rather than for quantification of normal Hb or thalassemia diagnosis. There are many reports showing the agreement of results obtained from HPLC and those obtained from other techniques such as the globin synthesis technique, isoelectrofocusing, carboxymethylcellulose chromatography and DNA sequencing , Rao et al, 1997, Fucharoen et al, 1998.…”

Section: High Performance Liquid Chromatographymentioning

confidence: 82%

“…High-performance liquid chromatographic (HPLC) methods with high sensitivity and specificity have been developed for both screening and confirmation of hemoglobinopathies and thalassemia in newborns. The HPLC technique requires a very small amount of blood samples (µl), therefore, it is very suitable for prenatal diagnosis of thalassemia (Maiavacca et al, 1992, Rao et al, 1997, where sample may be limited and difficult to obtain. In HPLC, particle size of the stationary phase packed in the column is quite small (about 2-5µm).…”

Section: High Performance Liquid Chromatographymentioning

confidence: 99%

Thalassemia Syndrome

Tangvarasittichai¹

2011

Advances in the Study of Genetic Disorders

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“…One of the limitations of fetal blood analysis by this analyzer is that it cannot distinguish between β + /β 0 and β + /β + homozygotes and between β + heterozygotes and normal subjects. In 1997, Rao et al (11) showed the Hb A level for 27 affected fetuses to be 0%, while two showed a Hb A value of 0.5%. The mean Hb A for 61 unaffected fetuses was 4.8 ± 2.08%.…”

Section: Discussionmentioning

confidence: 99%

Is the Poly A (T>C) Mutation a Causative Factor For Misdiagnosis in Second Trimester Prenatal Diagnosis of β-Thalassemia by Fetal Blood Analysis on High Performance Liquid Chromatography?

et al. 2012

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We report the problems in diagnosis faced by two families referred for prenatal diagnosis of thalassemia where cordocentesis and fetal blood analysis by high performance liquid chromatography (HPLC) had to be done. The Hb A levels of the fetal blood measured by HPLC on the VARIANT™ Hemoglobin Testing System were 1.2 and 6.7%, respectively, suggestive of a heterozygous β-thalassemia (β-thal) fetus in the first case and a normal fetus in the second case. In one family, one of the parents had a borderline Hb A(2) level and in the other, one parent had normal RBC indices. However, DNA sequencing, done later, showed that in the first case the fetus was a compound heterozygote for the IVS-I-5 (G>C) and the polyadenylation signal site [poly A (T>C)] mutation, while in the second case, the fetus was homozygous for the poly A mutation. This emphasizes that characterization of β-thal mutations must be done whenever one of the parents has a borderline Hb A(2) level or normal RBC indices, and one should not rely on fetal blood analysis by HPLC for prenatal diagnosis of β-thal so as to avoid misdiagnosis.

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Rapid Mid-Trimester Prenatal Diagnosis of Beta-thalassaemia and other Haemoglobinopathies using a Non-Radioactive Anion Exchange HPLC Technique—An Indian Experience

Cited by 14 publications

References 14 publications

Analysis of fetal blood using capillary electrophoresis system: a simple method for prenatal diagnosis of severe thalassemia diseases

Analysis of fetal blood using capillary electrophoresis system: a simple method for prenatal diagnosis of severe thalassemia diseases

Thalassemia Syndrome

Is the Poly A (T>C) Mutation a Causative Factor For Misdiagnosis in Second Trimester Prenatal Diagnosis of β-Thalassemia by Fetal Blood Analysis on High Performance Liquid Chromatography?

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