Hb Rush (HBB: c.304G>C): A Rare Variant Hemoglobin Mimicking the Hb S (HBB: c.20A>T) Variant on High Performance Liquid Chromatography

Hemoglobin ◽  
2020 ◽  
Vol 44 (1) ◽  
pp. 64-66
Author(s):  
Ranjeet S. Mashon ◽  
Reena Das ◽  
Radhika Dhawan ◽  
Naveen Kakkar ◽  
Tapasya Dhar
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Rare Variant ◽  
High Performance ◽  
Variant Hemoglobin ◽  
Hb S

scholarly journals Study of High Performance Liquid Chromatography and Turbidimetric Inhibition Immunoassay for HbA1c Analysis in Diabetic Patients with Variant Hemoglobin

2018 ◽  
Vol 8 (2) ◽  
pp. 114-117
Author(s):  
Gazi Sharmin Sultana ◽  
Nadia Zebin Khan ◽  
Zannat E Khuda ◽  
Tanvira Afroze Sultana ◽  
Tashmim Farhana Dipta ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Glycemic Control ◽  
High Performance ◽  
Hplc Method ◽  
Diabetic Patients ◽  
Genetic Changes ◽  
Variant Hemoglobin ◽  
Hemoglobin Variants ◽  
Hba1c Levels

Background: HbA1c is considered as “gold standard” to evaluate glycemic control in patients with diabetes. Hemoglobin variants are mutant forms of hemoglobin that can occur by genetic changes in specific amino acid that can affect the accuracy of HbA1c measurements. High performance liquid chromatography (HPLC) is the standard method for HbA1c but inaccurate HbA1c values can occur when hemoglobin variants are present in diabetic patient. The aim of our study is to see Turbidimetric Inhibition Immunoassay (TINIA) method can report HbA1c values in diabetic patients with variant hemoglobin when the values are inaccurate on HPLC.Methods: 7590 diabetic patients were analyzed for HbA1c by HPLC method from BIRDEM General Hospital during December 2013 to January 2014. HbA1c levels were again measured by TINIA method in 50 cases out of 7590 who showed either undetectable / below normal HbA1c levels. Hb electrophoresis confirmed the variant hemoglobin in few casesResults: 50 cases out of 7590 (0.65%) had either undetectable / below normal HbA1c levels by HPLC method. Males-26 and females-24; and the ratio was 0.92:1. In 27 cases, HbA1c values were undetectable by HPLC method but in the reportable range by TINIA method. In the other 23 cases, HbA1c levels were below the reportable range (<4%) by HPLC method but were in the normal or higher range by TINIA method. On Bland Altman plot, TINIA method did not agree with HPLC method in variant cases.Conclusion: In South East Asia where Hb variant is high, Low or undetectable HbA1c level by HPLC may be a convenient clue for screening of hemoglobinopathies especially among diabetic population in Bangladesh. All laboratories should have alternative method of HbA1c testing like TINIA along with HPLC for correct determination of glycemic control in variant casesBirdem Med J 2018; 8(2): 114-117

Unintended Reporting of Misleading Hb A1c Values When Using Assays Incapable of Detecting Hemoglobin Variants

2013 ◽  
Vol 137 (12) ◽  
pp. 1788-1791 ◽  
Author(s):  
Jeanne M. Rhea ◽  
David Koch ◽  
James Ritchie ◽  
Harsh V. Singh ◽  
Andrew N. Young ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Compound Heterozygous ◽  
Hb A1c ◽  
Cell Life ◽  
High Prevalence ◽  
Hb S ◽  
Hemoglobin Variants ◽  
Hb Variant

Context.—It is recommended that hemoglobin (Hb) A1c (Hb A1c) not be used to assess average glycemia in patients who have altered red blood cell life span. Objective.—To investigate the frequency of reporting an Hb A1c value for Hb variant samples that do not include Hb A. Design.—Hb A1c samples (n = 500) were procured and screened for Hb variants that may affect Hb A1c interpretation (Hb SS, Hb SC, and Hb S–β-thalassemia). Five of each of these samples were tested by ion-exchange high-performance liquid chromatography, immunoturbidimetric assay, second-generation immunoturbidimetric assay, and affinity chromatography. Results.—Eleven (2.2%) homozygous Hb SS, 6 (1.2%) Hb SC, and 5 (1.0%) Hb S–β-thalassemia samples were identified out of the 500 samples tested. Three of 4 instruments investigated in this study are known to not be plagued by analytic interference from these Hb variants but disturbingly reported Hb A1c values in the absence of Hb A. Conclusions.—The improved analytic specificity of Hb A1c platforms has by and large eliminated interferences from the most common heterozygous Hb variants. A consequence, however, is the potential for unintended reporting of Hb A1c results in the presence of homozygous and compound heterozygous Hb variants that lack Hb A and the inability to distinguish those samples not recommended to be used for patient care. The ability to identify samples harboring Hb variants that preclude the utility of Hb A1c may be beneficial in high prevalence populations.

Quantification of hemoglobins S, C, and F by a magnetic affinity immunoassay.

1988 ◽  
Vol 34 (5) ◽  
pp. 902-905 ◽  
Author(s):  
H Moscoso ◽  
C R Kiefer ◽  
A Kutlar ◽  
F A Garver
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Enzyme Immunoassay ◽  
High Performance ◽  
Magnetic Beads ◽  
Percent Inhibition ◽  
Standard Curve ◽  
Hb S ◽  
Monospecific Antisera ◽  
Hb C

Abstract This magnetic affinity immunoassay (MAIA) quantifies hemoglobins (Hb) S, C, and F in hemolysates from adults or newborns. Monospecific antisera to the hemoglobins are covalently conjugated to magnetic beads and reacted with the corresponding 125I-labeled hemoglobin. After centrifugation to separate the free and antibody-bound 125I-labeled hemoglobin, the amount of radioactive hemoglobin in the pellet is measured. To determine the concentration of the Hb under study, the percent inhibition of the reaction is quantified. The standard curve is established by adding known quantities of unlabeled hemoglobin before adding 125I-labeled hemoglobin. The amount of Hb S, Hb C, or Hb F present in hemolysates is determined by measuring the percentage of inhibition and extrapolating the concentration from the standard curve. Results agree well with values for Hb S and Hb C obtained by "high-performance" liquid chromatography and RIA and for Hb F as measured by alkali denaturation and RIA. This assay can be completed in 1 h and is more sensitive than enzyme immunoassay or RIA: we can detect proportions of Hb S and Hb C as low as 1% of total Hb, and Hb F as low as 0.05%.

Laboratory Recognition of a Rare Hemoglobinopathy

1999 ◽  
Vol 123 (10) ◽  
pp. 963-966
Author(s):  
Cheryl M. Kirk ◽  
Christine N. Papadea ◽  
John Lazarchick
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Dna Analysis ◽  
Clinical Course ◽  
Chain Reaction ◽  
Molecular Defects ◽  
Alpha Globin ◽  
Hb S ◽  
Polymerase Chain

Abstract This article describes the laboratory investigation of an unusual hemoglobinopathy involving hemoglobin (Hb) S, HbSGPhiladelphia, and α-thalassemia-2 in a patient whose phenotype was HbSC by alkaline electrophoresis. Findings of a mean corpuscular volume of 62 fL and microcytes on the blood smear were inconsistent with HbSC disease. The patient's clinical course over several years had been mildly symptomatic. Testing in our hospital laboratory using isoelectric focusing and cation-exchange high-performance liquid chromatography to separate hemoglobins showed an unknown variant. Additional studies, including globin chain electrophoresis, reverse-phase high-performance liquid chromatography, and polymerase chain reaction–based DNA analysis were performed at reference laboratories, which reported the following findings: HbGPhiladelphia associated with α-thalassemia-2, HbS and HbGPhiladelphia, and the alpha-globin deletions defining the −α3.7/−α3.7 genotype. The hemoglobin molecular defects, α-thalassemia-2, and the pattern of inheritance are discussed.

scholarly journals Accidental detection of clinically silent compound heterozygous Hb D Punjab/Hb Q India while analyzing HbA1c by high performance liquid chromatography

2021 ◽  
Vol 9 (11) ◽  
pp. 3468
Author(s):  
Hiren J. Dhanani ◽  
Mittal C. Sukhadiya ◽  
Nandini H. Dhanani ◽  
Jaysukh D. Kothia ◽  
Bharart D. Tandel
Keyword(s):  
High Performance Liquid Chromatography ◽  
Capillary Electrophoresis ◽  
Liquid Chromatography ◽  
High Performance ◽  
Compound Heterozygous ◽  
Hemoglobin Variant ◽  
Variant Hemoglobin ◽  
Hbq India ◽  
Second Peak ◽  
Cation Exchange Hplc

HbA1c is routinely prescribed investigations for diagnosing and monitoring diabetes and high-performance liquid chromatography (HPLC) is preferred method which is also able to identify presence of hemoglobin variant. A case was encountered where presence of variant hemoglobin was indicated. On further investigation with three different instruments, diagnosis of compound heterozygous Hb D Punjab/Hb Q India was made. The chromatogram on Bio-Rad D10 showed Hb D Punjab (ααββHbD Punjab)-29.89% at 3.96 minutes retention time (RT), Hb Q India (ααHbQ Indiaββ) -9.5% with 4.45 minutes RT, hybrid of HbQ India/Hb D Punjab (ααHbQ IndiaββHbD Punjab)-6% with 4.66 minutes RT, Hb A2 (ααδδ) was 2.5% and Hb A (ααββ) was 52.2%. Analysis done on Bio-Rad variant V-II confirmed these findings. Analysis done on Sebia capillary electrophoresis revealed major peak of 50.9% in zone 9/Z(A) constituted by Hb A, second peak of 39.8% in zone 6/Z(D) constituted by co-elution of Hb D and Hb Q India, third peak of 8.8% in zone 3-4/Z(A2-C) constituted by co-elution of Hb A2 and hybrid of Hb D Punjab/Hb Q India and a fourth peak of 0.5% in zone 1 representing Hb A2HbQ India (ααHbQ Indiaδδ). Ideally variants detected while analyzing HbA1c should be further investigated for confirmation and result of which should be shared, discussed and the patient should be encouraged for screening of available family members for relevant variant hemoglobin. Combination of cation exchange HPLC and capillary electrophoresis in this case was sufficient to arrive at conclusion.

scholarly journals Comparing electrophoresis at alkaline pH and high performance liquid chromatography to diagnose Hb S-like hemoglobin

2012 ◽  
Vol 34 (2) ◽  
pp. 170-171
Author(s):  
Renan Gracia de Oliveira ◽  
Paula Juliana Antoniazzo Zamaro ◽  
Claudia Regina Bonini-Domingos
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Alkaline Ph ◽  
Hb S
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