Rapid molecular diagnostics of large deletional β0-thalassemia (3.5 kb and 45 kb) using colorimetric LAMP in various thalassemia genotypes

Background: β-thalassemia is an inherited disorder that is reported worldwide. Two common β0-thalassemia mutations (3.5 kb and 45 kb deletions) are prevalent in Southeast Asia and Thailand. Identification of these defects is essential to population screening and prenatal diagnosis. We aimed to devel...

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Autores principales:	Wanicha Tepakhan, Wittaya Jomoui
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	3.5 kb deletion 45 kb deletion β-thalassemia Loop-mediated isothermal amplification Molecular testing Science (General) Q1-390 Social sciences (General) H1-99
Acceso en línea:	https://doaj.org/article/8aec7bcf5ada485ba55aa0cad2b54634
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spelling	oai:doaj.org-article:8aec7bcf5ada485ba55aa0cad2b546342021-12-02T05:02:53ZRapid molecular diagnostics of large deletional β0-thalassemia (3.5 kb and 45 kb) using colorimetric LAMP in various thalassemia genotypes2405-844010.1016/j.heliyon.2021.e08372https://doaj.org/article/8aec7bcf5ada485ba55aa0cad2b546342021-11-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2405844021024750https://doaj.org/toc/2405-8440Background: β-thalassemia is an inherited disorder that is reported worldwide. Two common β0-thalassemia mutations (3.5 kb and 45 kb deletions) are prevalent in Southeast Asia and Thailand. Identification of these defects is essential to population screening and prenatal diagnosis. We aimed to develop colorimetric LAMP based on a phenol red indicator and validate it on various thalassemia genotypes. Method: Colorimetric LAMP assays for detecting β0-thalassemia 3.5- and 45-kb deletions were developed and validated on 254 routine clinical samples. The results of the assays could be interpreted by the naked eye and compared with the gold standard gap-PCR. Results: A total of 254 samples related to seven phenotypes and 27 different genotype groups showed 100% concordance between the colorimetric LAMP assays and gap-PCR for detecting β0-thalassemia (3.5- and 45-kb deletions). The sensitivity, specificity, NPV, and PPV were calculated as 100% for both β0-thalassemia 3.5- and 45-kb deletion detection. The comparison of the usefulness of colorimetric LAMP assays and conventional methods was demonstrated in this study. Conclusions: The developed colorimetric LAMP assays are rapid, simple, and highly cost effective and can be interpreted by the naked eye. These assays should be applied for screening deletional β0-thalassemia in routine settings or small community hospitals in remote areas where thalassemia is highly heterogeneous.Wanicha TepakhanWittaya JomouiElsevierarticle3.5 kb deletion45 kb deletionβ-thalassemiaLoop-mediated isothermal amplificationMolecular testingScience (General)Q1-390Social sciences (General)H1-99ENHeliyon, Vol 7, Iss 11, Pp e08372- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	3.5 kb deletion 45 kb deletion β-thalassemia Loop-mediated isothermal amplification Molecular testing Science (General) Q1-390 Social sciences (General) H1-99
spellingShingle	3.5 kb deletion 45 kb deletion β-thalassemia Loop-mediated isothermal amplification Molecular testing Science (General) Q1-390 Social sciences (General) H1-99 Wanicha Tepakhan Wittaya Jomoui Rapid molecular diagnostics of large deletional β0-thalassemia (3.5 kb and 45 kb) using colorimetric LAMP in various thalassemia genotypes
description	Background: β-thalassemia is an inherited disorder that is reported worldwide. Two common β0-thalassemia mutations (3.5 kb and 45 kb deletions) are prevalent in Southeast Asia and Thailand. Identification of these defects is essential to population screening and prenatal diagnosis. We aimed to develop colorimetric LAMP based on a phenol red indicator and validate it on various thalassemia genotypes. Method: Colorimetric LAMP assays for detecting β0-thalassemia 3.5- and 45-kb deletions were developed and validated on 254 routine clinical samples. The results of the assays could be interpreted by the naked eye and compared with the gold standard gap-PCR. Results: A total of 254 samples related to seven phenotypes and 27 different genotype groups showed 100% concordance between the colorimetric LAMP assays and gap-PCR for detecting β0-thalassemia (3.5- and 45-kb deletions). The sensitivity, specificity, NPV, and PPV were calculated as 100% for both β0-thalassemia 3.5- and 45-kb deletion detection. The comparison of the usefulness of colorimetric LAMP assays and conventional methods was demonstrated in this study. Conclusions: The developed colorimetric LAMP assays are rapid, simple, and highly cost effective and can be interpreted by the naked eye. These assays should be applied for screening deletional β0-thalassemia in routine settings or small community hospitals in remote areas where thalassemia is highly heterogeneous.
format	article
author	Wanicha Tepakhan Wittaya Jomoui
author_facet	Wanicha Tepakhan Wittaya Jomoui
author_sort	Wanicha Tepakhan
title	Rapid molecular diagnostics of large deletional β0-thalassemia (3.5 kb and 45 kb) using colorimetric LAMP in various thalassemia genotypes
title_short	Rapid molecular diagnostics of large deletional β0-thalassemia (3.5 kb and 45 kb) using colorimetric LAMP in various thalassemia genotypes
title_full	Rapid molecular diagnostics of large deletional β0-thalassemia (3.5 kb and 45 kb) using colorimetric LAMP in various thalassemia genotypes
title_fullStr	Rapid molecular diagnostics of large deletional β0-thalassemia (3.5 kb and 45 kb) using colorimetric LAMP in various thalassemia genotypes
title_full_unstemmed	Rapid molecular diagnostics of large deletional β0-thalassemia (3.5 kb and 45 kb) using colorimetric LAMP in various thalassemia genotypes
title_sort	rapid molecular diagnostics of large deletional β0-thalassemia (3.5 kb and 45 kb) using colorimetric lamp in various thalassemia genotypes
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/8aec7bcf5ada485ba55aa0cad2b54634
work_keys_str_mv	AT wanichatepakhan rapidmoleculardiagnosticsoflargedeletionalb0thalassemia35kband45kbusingcolorimetriclampinvariousthalassemiagenotypes AT wittayajomoui rapidmoleculardiagnosticsoflargedeletionalb0thalassemia35kband45kbusingcolorimetriclampinvariousthalassemiagenotypes
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Rapid molecular diagnostics of large deletional β0-thalassemia (3.5 kb and 45 kb) using colorimetric LAMP in various thalassemia genotypes

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