HPLC studies in hemoglobinopathies

Abstract

An accurate diagnosis of beta -thalassemia carriers, homozygous patients and identification of different structural hemoglobin variants is important for epidemiological studies as well as for management and prevention of the major hemoglobin disorders. There are many electrophoretic and chromatographic approaches for estimation of HbA2 and Hb F but cation exchange HPLC (CE-HPLC)using automated dedicated machines like the Variant Hb testing system have become the method of choice for these investigations. CE-HPLC also helps in the presumptive identification of many abnormal hemoglobin variants and has been useful for both neonatal screening of sickle cell disease as well as second trimester prenatal diagnosis of thalassemia by fetal blood analysis. Other applications of HPLC in hemoglobinopathies include separation of globin chains, measuring the ratio of gamma globin chains (Ggamma/Agamma) and the recently described denaturing HPLC for detecting mutations in both alpha and beta globin genes.

Frequency of haemoglobinopathies: a single-centre, cross-sectional study from Islamabad, Pakistan

“…A majority of the haemoglobinopathies are not clinically apparent but some produce serious life-threatening diseases and constitute a significant health care burden. These are quantitative (thalassaemia syndromes) or qualitative (variant Hb) [4][5][6][7]. Thalassaemia syndromes are subclassified based on the gene involved, i.e.…”

O laboratório clínico na investigação dos distúrbios da hemoglobina

“…A maioria dos programas de triagem populacional utiliza o valor de VCM < 80 fL e uma HCM < 27 pg como pontos de corte para a investigação adicional das talassemias (7) . Esses dados, em um contexto de RBC normal/alto, são altamente sugestivos para o diagnóstico de talassemia, como observado no quarto caso.…”

Hemoglobin J—As a Cause of Congenital Hemolytic Anemia

“…Today, CE-HPLC has become the method of choice for quantification of Hb A2, Hb F and other Hb subtypes [9,10]. A large prospective study was undertaken in the United States of America on over 60,000 samples analyzed by CE-HPLC on the Variant II machine over a 32 months period in a multiethnic population [10].…”

“…The authors, through this case, emphasize the need to utilize HPLC studies to detect Hb variants in addition to the usual Hb electrophoresis. Also, CE-HPLC along with isoelectric focusing has become the method of choice for newborn and prenatal screening of hemoglobinopathies [9].…”