Capillary zone electrophoresis for haemoglobinopathy diagnosis

Borbely, Nicole; Phelan, Lorraine; Szydlo, Richard; Bain, Barbara J.

doi:10.1136/jclinpath-2012-200946

Cited by 41 publications

(23 citation statements)

References 9 publications

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“…Traditional method of β-thalassemia screening including hematological parameters and hemoglobin electrophoresis could not be applied for dry blood spots of neonates. Some techniques could be used for screening β-thalassemia by dry blood spots, such as high performance liquid chromatography (HPLC) and capillary electrophoresis (CE) [20], [21], and amplification refractory mutation system-PCR, PCR-SSCP, denaturing high performance liquid chromatography (DHPLC), matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF) and PCR-RDB [6]. However, these methods are time consuming and costly for large scale screening.…”

Section: Discussionmentioning

confidence: 99%

High Resolution Melting Analysis: A Rapid Screening and Typing Tool for Common β-Thalassemia Mutation in Chinese Population

et al. 2014

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β-thalassemia is a common inherited disorder worldwide including southern China, and at least 45 distinct β-thalassemia mutations have been identified in China. High-resolution melting (HRM) assay was recently introduced as a rapid, inexpensive and effective method for genotyping. However, there was no systemic study on the diagnostic capability of HRM to identify β-thalassemia. Here, we used an improved HRM method to screen and type 12 common β-thalassemia mutations in Chinese, and the rapidity and reliability of this method was investigated. The whole PCR and HRM procedure could be completed in 40 min. The heterozygous mutations and 4 kinds of homozygous mutations could be readily differentiated from the melting curve except c.-78A>G heterozygote and c.-79A>G heterozygote. The diagnostic reliability of this HRM assay was evaluated on 756 pre-typed genomic DNA samples and 50 cases of blood spots on filter paper, which were collected from seven high prevalent provinces in southern China. If c.-78A>G heterozygote and c.-79A>G heterozygote were classified into the same group (c.-78&79 A>G heterozygote), the HRM method was in complete concordance with the reference method (reverse dot blot/DNA-sequencing). In a conclusion, the HRM method appears to be an accurate and sensitive method for the rapid screening and identification of β-thalassemia mutations. In the future, we suggest this technology to be used in neonatal blood spot screening program. It could enlarge the coverage of β-thalassemia screening program in China. At the same time, its value should be confirmed in prospectively clinical and epidemiological studies.

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Section: Discussionmentioning

confidence: 99%

High Resolution Melting Analysis: A Rapid Screening and Typing Tool for Common β-Thalassemia Mutation in Chinese Population

et al. 2014

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“…In addition to chronicling elution times for identification of the hemoglobin variant, HPLC also provides percent compositions of blood samples. CE can also provide a quantitative evaluation of hemoglobin types [10,11]. …”

Section: Discussionmentioning

confidence: 99%

First North American case of Hemoglobin Shepherds Bush (β 74[E18] Gly → Asp) in a central Pennsylvania family

et al. 2014

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BackgroundHemoglobin Shepherds Bush (Human Genome Variation Society name: HBB:c.224G > A) is an unstable hemoglobin variant resulting from a β 74 GGC to GAC mutation (Gly to Asp) that manifests clinically as hemolytic anemia or gall bladder disease due to chronic subclinical hemolysis.Case presentationWe report a Pennsylvania family of English descent with this condition, first noticed in a 6-year-old female. The proband presented with splenomegaly, fatigue, dark urine and an elevated indirect bilirubin. Hemoglobin identification studies and subsequent genetic testing performed according to a systematic algorithm elucidated the diagnosis of Hb Shepherds Bush.ConclusionsThis is the first case of this rare hemoglobin variant identified in North America to our knowledge. It was identified using a systematic algorithm of diagnostic tests that should be followed whenever considering a rare hemoglobinopathy as part of the differential diagnosis.

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“…It is not prone to common interferences and has the ability to detect several hemoglobin variants. 9 Borbely and collegues 10 have observed some uncommon variant hemoglobins that are detected by capillary electrophoresis but not HPLC, although the reverse also occurs in some cases. …”

Section: Figurementioning

confidence: 99%

Interference of the Hope Hemoglobin With Hemoglobin A1c Results

Chakraborty

Chanda

Gain

et al. 2015

Lab Med

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Hemoglobin A1c (HbA1c) is now considered to be the marker of choice in diagnosis and management of diabetes mellitus, based on the results of certain landmark clinical trials. Herein, we report the case of a 52-year-old ethnic Southeast Asian Indian man with impaired glucose tolerance whose glycated hemoglobin (ie, HbA1c) levels, as measured via Bio-Rad D10 high-performance liquid chromatography (HPLC) and Roche Tina-quant immunoassay were 47.8% and 44.0%, respectively. No variant hemoglobin (Hb) peak was observed via the D10 chromatogram. We assayed the patient specimen on the Sebia MINICAP capillary electrophoresis platform; the HbA1c level was 6.8%, with a large variant Hb peak of 42.0%. This finding suggested the possible presence of the heterozygous Hb Hope, which can result in spuriously elevated HbA1c results on HPLC and turbidimetric immunoassays. Although the capillary electrophoresis system was able to identify the variant, the A1c results should not be considered accurate due to overlapping of the variant and adult Hb peaks on the electrophoretogram reading. Hb Hope is usually clinically silent but can present such analytical challenges. Through this case study, we critically discuss the limitations of various HbA1c assay methods, highlighting the fact that laboratory professionals need to be aware of occurrences of Hb Hope, to help ensure patient safety.

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Capillary zone electrophoresis for haemoglobinopathy diagnosis

Abstract: Capillary electrophoresis using a Capillarys 2 Flex Piercing instrument is suitable for haemoglobinopathy diagnosis.

Cited by 41 publications

References 9 publications

High Resolution Melting Analysis: A Rapid Screening and Typing Tool for Common β-Thalassemia Mutation in Chinese Population

High Resolution Melting Analysis: A Rapid Screening and Typing Tool for Common β-Thalassemia Mutation in Chinese Population

First North American case of Hemoglobin Shepherds Bush (β 74[E18] Gly → Asp) in a central Pennsylvania family

Interference of the Hope Hemoglobin With Hemoglobin A1c Results

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