Screening of (–<sup>SEA</sup>) α-thalassaemia using an immunochromatographic strip assay for the ζ-globin chain in a population with a high prevalence and heterogeneity of haemoglobinopathies

Jomoui, Wittaya; Fucharoen, Goonnapa; Sanchaisuriya, Kanokwan; Fucharoen, Supan

doi:10.1136/jclinpath-2016-203765

Cited by 10 publications

(8 citation statements)

References 23 publications

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“…It is noted that within subjects carrying non-α thalassemia 1 gene, by the immunostick test, approximately 20% of these subjects showed positive reactivity. These results are similar to those obtained previously by using other immunoassay methods [17, 18]. The cause of this positivity, however, is still unknown.…”

Section: Discussionsupporting

confidence: 91%

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Immunostick Test for Detecting ζ-Globin Chains and Screening of the Southeast Asian α-Thalassemia 1 Deletion

et al. 2019

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Background Couples who carry α-thalassemia-1 deletion are at 25% risk of having a fetus with hemoglobin Bart’s hydrops fetalis. Southeast Asian deletion (−-(SEA)) is the most common type of α-thalassemia 1 among Southeast Asian populations. Thus, identification of the (−-(SEA)) α-thalassemia 1 carrier is necessary for controlling severe α-thalassemia in Southeast Asian countries. Results Using our generated anti ζ-globin chain monoclonal antibodies (mAbs) clones PL2 and PL3, a simple immunostick test for detecting ζ-globin chain presence in whole blood lysates was developed. The procedure of the developed immunostick test was as follows. The immunostick paddles were coated with 50 μg/mL of mAb PL2 as capture mAb, or other control antibodies. The coated immunostick was dipped into cocktail containing tested hemolysate at dilution of 1:500, 0.25 μg/mL biotin-labeled mAb PL3 and horseradish peroxidase-conjugated streptavidin at dilution of 1:1000. The immunostick was then dipped in precipitating substrate and the presence of ζ-globin chain in the tested sample was observed by the naked eye. Upon validation of the developed immunostick test with various types of thalassemia and normal subjects, 100% sensitivity and 82% specificity for detection of the (−-(SEA)) α-thalassemia-1 carriers were achieved. The mAb pre-coated immunostick can be stored at room temperature for at least 20 weeks. Conclusion In this study, a novel simple immunostick test for the screening of (−-(SEA)) α-thalassemia 1 carriers was presented. The developed immunostick test, within a single test, contains both positive and negative internal procedural controls.

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Section: Discussionsupporting

confidence: 91%

“…The cause of this positivity, however, is still unknown. The cross-reactivity of anti-ζ-globin chain mAbs used with other globin chains have been assumed [18].…”

Section: Discussionmentioning

confidence: 99%

Immunostick Test for Detecting ζ-Globin Chains and Screening of the Southeast Asian α-Thalassemia 1 Deletion

et al. 2019

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“…These results are similar to those obtained previously [21, 25–27, 35, 36]. The cause of the positivity of the IC strip tests with these subjects is still unknown and may be due to cross-reactivity of mAbs used in the IC strip tests to other Hbs [35]. Although the IC strip tests could detect other thalassemias in addition to α 0 -thalassemia, this test will have a great benefit for the identification of β-thalassemia subjects that also have α-thalassemia genes [26].…”

Section: Discussionsupporting

confidence: 93%

“…All α 0 -thalassemia (-- SEA ) samples tested positive using the IC ζ strip test. These results were in accordance with previous reports showing that small amounts of embryonic ζ-globin chains are present in hemolysates and can serve as a marker for (-- SEA ) α 0 -thalassemia traits [32, 35, 36]. Accordingly, the negative IC ζ strip test results could exclude α 0 -thalassemia (-- SEA ) carriers.…”

Section: Discussionsupporting

confidence: 92%

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Impact of the detection of ζ-globin chains and hemoglobin Bart’s using immunochromatographic strip tests for α0-thalassemia (--SEA) differential diagnosis

et al. 2019

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α0-Thalassemia is an inherited hematological disorder caused by the deletion of α-globin genes. The Southeast Asian deletion (--SEA) is the most common type of α0-thalassemia observed in Southeast Asian countries. Regarding WHO health policy, an effective α0-thalassemia screening strategy is needed to control new severe α-thalassemia cases. In this study, a monoclonal antibody panel was used to develop immunochromatographic (IC) strip tests for detecting the Hb Bart’s and ζ-globin chain. Among 195 samples, all α0-thalassemia traits (78 α0-thalassemia (--SEA) and 4 α0-thalassemia (--THAI)) had low MCV or MCH values. The sensitivity, specificity, PPV and NPV of the IC strip tests for ζ-globin and Hb Bart’s for screening α0-thalassemia (--SEA) within the low MCV or MCH samples were 100%, 65.2%, 90.7%, 100% and 96.2%, 47.8%, 86.6%, 78.6%, respectively. All 4 α0-thalassemia (--THAI) traits were negative for ζ-globin chains but positive for Hb Bart’s using the IC strip tests. These results led to a α0-thalassemia screening being proposed in which blood samples are first evaluated by MCV, MCH and Hb typing. Samples with high MCV and MCH values are excluded for the presence of the α0-thalassemia gene. Samples with low MCV or MCH values are assayed using the developed IC strip tests, where only samples testing positive are further assayed for α0-thalassemia by PCR. Patients with Hb H, EA Bart’s or EF Bart’s diseases do not need to use this IC strip assay. Thus, in this study, a simple and cost effective α0-thalassemia point of care test was developed.

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Laboratory techniques for the identification of abnormalities of globin chain synthesis

2020

Haemoglobinopathy Diagnosis

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Screening of (–^SEA) α-thalassaemia using an immunochromatographic strip assay for the ζ-globin chain in a population with a high prevalence and heterogeneity of haemoglobinopathies

Cited by 10 publications

References 23 publications

Immunostick Test for Detecting ζ-Globin Chains and Screening of the Southeast Asian α-Thalassemia 1 Deletion

Immunostick Test for Detecting ζ-Globin Chains and Screening of the Southeast Asian α-Thalassemia 1 Deletion

Impact of the detection of ζ-globin chains and hemoglobin Bart’s using immunochromatographic strip tests for α0-thalassemia (--SEA) differential diagnosis

Laboratory techniques for the identification of abnormalities of globin chain synthesis

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Screening of (–SEA) α-thalassaemia using an immunochromatographic strip assay for the ζ-globin chain in a population with a high prevalence and heterogeneity of haemoglobinopathies

Cited by 10 publications

References 23 publications

Immunostick Test for Detecting ζ-Globin Chains and Screening of the Southeast Asian α-Thalassemia 1 Deletion

Immunostick Test for Detecting ζ-Globin Chains and Screening of the Southeast Asian α-Thalassemia 1 Deletion

Impact of the detection of ζ-globin chains and hemoglobin Bart’s using immunochromatographic strip tests for α0-thalassemia (--SEA) differential diagnosis

Laboratory techniques for the identification of abnormalities of globin chain synthesis

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Screening of (–^SEA) α-thalassaemia using an immunochromatographic strip assay for the ζ-globin chain in a population with a high prevalence and heterogeneity of haemoglobinopathies