SCREENING AND MOLECULAR CHARACTERIZATION OF HAEMOGLOBINOPATHIES USING ARMS (AMPLIFICATION REFRACTORY MUTATION SYSTEM) AND DIRECT DNA SEQUENCING TECHNIQUES AMONG YOUNG IN CENTRAL GUJARAT WESTERN INDIA

Haemoglobinopathies is consider the most common inherited disorders in human and results from genetic mutation in . one or more genes The present study was aimed to characterize the β-thalassemia mutation and haemoglobin variant in youth by ARMS PCR which is an uncomplicated and convenient method for identification of five common mutation from central Gujarat, western India region. This Study included 44 randomly selected haemoglobinopathies carrier student's sample of Anand People's Medicare Society (APMS), Anand for DNA analysis by ARMS PCR from March 2021-April 2021. Identification of five common Indian β thalassemia mutations along with Hb S and HB E were carried out by ARMS PCR method and δβ- thalassemia mutation was characterized by GAP-PCR. The samples which remain uncharacterized were sent to S N gene lab, Surat for DNA sequencing. In our study the most common mutation among five common mutations characterized was IVS-1, nt5 (G→C) in 22 (50%), followed by Codon41/42 (-CTTT) in 5 (11.3%). IVS-1, nt1 (G→T), Codon8/9 (+G) and 619bp del mutation was not identified in any carrier students screened for haemoglobinopathies. Other than these five common mutation Codon -88 (C→T) (2.27%) and Codon 30 (G→A) (2.27%) are also detected. The prevalence of haemoglobinopathies with respect to communities, reflects that SC/ST/OBC are at the highest risk with 50%. Communities like Rajput (22.7%), Patel (18.1%), Brahmin (6.8%) and Muslim (2.2%) are also showing prevalence. The study has included mutation in different communities reflects characterization of mutation of central Gujarat, western India which is significant for rapid and convenient identification of mutation while conducting screening programs and prenatal diagnosis. Rare mutations which are not recognized, need further confirmation for carrier detection followed by prenatal diagnosis.

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Molecular characterization of G6PD mutations reveals the high frequency of G6PD Aures in the Lao Theung population

10.21203/rs.3.rs-45493/v4 ◽

2020 ◽

Author(s):

Amkha Sanephonasa ◽

Chalisa Louicharoen Cheepsunthorn ◽

Naly Khaminsou ◽

Onekham Savongsy ◽

Issarang Nuchprayoon ◽

...

Keyword(s):

Enzyme Activity ◽

Ethnic Group ◽

Molecular Characterization ◽

G6pd Deficiency ◽

Gene Mutations ◽

Final Diagnosis ◽

Amplification Refractory Mutation System ◽

Common Mutation ◽

Arms Pcr

Abstract Background: The prevalence and genotypes of G6PD deficiency vary worldwide, with higher prevalence in malaria endemic areas. The first time assessment of G6PD deficiency prevalence and molecular characterization of G6PD mutations in the Lao Theung population were performed in this study. Methods: A total of 252 unrelated Lao Theung participants residing in the Lao People's Democratic Republic (PDR) were recruited. All participant samples were tested for G6PD enzyme activity and G6PD gene mutations. The amplification refractory mutation system (ARMS)-PCR for detecting G6PD Aures was developed.Results: The G6PD mutations were detected in 11.51% (29/252) of the participants. Eight G6PD mutations were detected. The G6PD Aures was the most common mutation identified in this cohort, which represented 58.62 % (17/29) of all mutation. The mutation pattern was homogenous, predominantly involving the G6PD Aures mutation (6.75%), followed by 1.19% G6PD Union and 0.79% each G6PD Jammu, G6PD Mahidol and G6PD Kaiping. One subject (0.4%) each carried G6PD Viangchan and G6PD Canton. Interestingly, one case of coinheritance of G6PD Aures and Quing Yan was detected in this cohort. Based on levels of G6PD enzyme activity, the prevalence of G6PD deficiency in the Lao Theung population was 9.13 % (23/252). The prevalence of G6PD deficient males and females (activity < 30 %) in the Lao Theung population was 6.41 % (5/78) and 1.72 % (3/174), respectively, and the prevalence of G6PD intermediate (activity 30-70 %) was 5.95 % (15/252).Conclusion: The G6PD Aures mutation is highly prevalent in the Lao Theung ethnic group. The common G6PD variants in continental Southeast Asian populations, G6PD Viangchan, Canton, Kaiping, Union and Mahidol, were not prevalent in this ethnic group. The technical simplicity of the developed ARMS-PCR will facilitate the final diagnosis of the G6PD Aures.

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Molecular characterization of G6PD mutations reveals the high frequency of G6PD Aures in the Lao Theung population

10.21203/rs.3.rs-45493/v5 ◽

2021 ◽

Author(s):

Amkha Sanephonasa ◽

Chalisa Louicharoen Cheepsunthorn ◽

Naly Khaminsou ◽

Onekham Savongsy ◽

Issarang Nuchprayoon ◽

...

Keyword(s):

Enzyme Activity ◽

Ethnic Group ◽

Molecular Characterization ◽

G6pd Deficiency ◽

Gene Mutations ◽

Final Diagnosis ◽

Amplification Refractory Mutation System ◽

Common Mutation ◽

Arms Pcr

Abstract Background The prevalence and genotypes of G6PD deficiency vary worldwide, with higher prevalence in malaria endemic areas. The first-time assessment of G6PD deficiency prevalence and molecular characterization of G6PD mutations in the Lao Theung population were performed in this study. Methods A total of 252 unrelated Lao Theung participants residing in the Lao People's Democratic Republic (PDR) were recruited. All participant samples were tested for G6PD enzyme activity and G6PD gene mutations. The amplification refractory mutation system (ARMS)-PCR for detecting G6PD Aures was developed.Results The G6PD mutations were detected in 11.51% (29/252) of the participants. Eight G6PD mutations were detected. The G6PD Aures was the most common mutation identified in this cohort, which represented 58.62 % (17/29) of all mutation. The mutation pattern was homogenous, predominantly involving the G6PD Aures mutation (6.75%), followed by 1.19% G6PD Union and 0.79% each G6PD Jammu, G6PD Mahidol and G6PD Kaiping. One subject (0.4%) each carried G6PD Viangchan and G6PD Canton. Interestingly, one case of coinheritance of G6PD Aures and Quing Yan was detected in this cohort. Based on levels of G6PD enzyme activity, the prevalence of G6PD deficiency in the Lao Theung population was 9.13 % (23/252). The prevalence of G6PD deficient males and females (activity < 30 %) in the Lao Theung population was 6.41 % (5/78) and 1.72 % (3/174), respectively, and the prevalence of G6PD intermediate (activity 30-70 %) was 5.95 % (15/252).Conclusion The G6PD Aures mutation is highly prevalent in the Lao Theung ethnic group. The common G6PD variants in continental Southeast Asian populations, G6PD Viangchan, Canton, Kaiping, Union and Mahidol, were not prevalent in this ethnic group. The technical simplicity of the developed ARMS-PCR will facilitate the final diagnosis of the G6PD Aures.

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Molecular characterization of G6PD mutations reveals the high frequency of G6PD Aures in the Lao Theung population

Malaria Journal ◽

10.1186/s12936-020-03560-7 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Amkha Sanephonasa ◽

Chalisa Louicharoen Cheepsunthorn ◽

Naly Khaminsou ◽

Onekham Savongsy ◽

Issarang Nuchprayoon ◽

...

Keyword(s):

Enzyme Activity ◽

Ethnic Group ◽

Molecular Characterization ◽

G6pd Deficiency ◽

Gene Mutations ◽

Final Diagnosis ◽

Amplification Refractory Mutation System ◽

Common Mutation ◽

Arms Pcr

Abstract Background The prevalence and genotypes of G6PD deficiency vary worldwide, with higher prevalence in malaria endemic areas. The first-time assessment of G6PD deficiency prevalence and molecular characterization of G6PD mutations in the Lao Theung population were performed in this study. Methods A total of 252 unrelated Lao Theung participants residing in the Lao People’s Democratic Republic (PDR) were recruited. All participant samples were tested for G6PD enzyme activity and G6PD gene mutations. The amplification refractory mutation system (ARMS)-PCR for detecting G6PD Aures was developed. Results The G6PD mutations were detected in 11.51% (29/252) of the participants. Eight G6PD mutations were detected. The G6PD Aures was the most common mutation identified in this cohort, which represented 58.62% (17/29) of all mutation. The mutation pattern was homogenous, predominantly involving the G6PD Aures mutation (6.75%), followed by 1.19% G6PD Union and 0.79% each G6PD Jammu, G6PD Mahidol and G6PD Kaiping. One subject (0.4%) each carried G6PD Viangchan and G6PD Canton. Interestingly, one case of coinheritance of G6PD Aures and Quing Yan was detected in this cohort. Based on levels of G6PD enzyme activity, the prevalence of G6PD deficiency in the Lao Theung population was 9.13% (23/252). The prevalence of G6PD deficient males and females (activity < 30%) in the Lao Theung population was 6.41% (5/78) and 1.72% (3/174), respectively, and the prevalence of G6PD intermediate (activity 30–70%) was 5.95% (15/252). Conclusions The G6PD Aures mutation is highly prevalent in the Lao Theung ethnic group. The common G6PD variants in continental Southeast Asian populations, G6PD Viangchan, Canton, Kaiping, Union and Mahidol, were not prevalent in this ethnic group. The technical simplicity of the developed ARMS-PCR will facilitate the final diagnosis of the G6PD Aures.

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Molecular characterization of G6PD mutations reveals the high frequency of G6PD Aures in the Lao Theung population

10.21203/rs.3.rs-45493/v2 ◽

2020 ◽

Author(s):

Amkha Sanephonasa ◽

Chalisa Louicharoen Cheepsunthorn ◽

Naly Khaminsou ◽

Onekham Savongsy ◽

Issarang Nuchprayoon ◽

...

Keyword(s):

Enzyme Activity ◽

Ethnic Group ◽

Molecular Characterization ◽

G6pd Deficiency ◽

Gene Mutations ◽

Final Diagnosis ◽

Amplification Refractory Mutation System ◽

Common Mutation ◽

Arms Pcr

Abstract Background: The prevalence and genotypes of G6PD deficiency vary worldwide, with higher prevalence in malaria endemic areas. The first time assessment of G6PD deficiency prevalence and molecular characterization of G6PD mutations in the Lao Theung population were performed in this study.Methods: A total of 252 unrelated Lao Theung participants residing in the Lao People's Democratic Republic (PDR) were recruited. All participant samples were tested for G6PD enzyme activity and G6PD gene mutations. The amplification refractory mutation system (ARMS)-PCR for detecting G6PD Aures was developed.Results: The G6PD mutations were detected in 11.51% (29/252) of the participants. Eight G6PD mutations were detected. The G6PD Aures was the most common mutation identified in this cohort, which represented 58.62 % (17/29) of all mutation. The mutation pattern was homogenous, predominantly involving the G6PD Aures mutation (6.75%), followed by 1.19% G6PD Union and 0.79% each G6PD Jammu, G6PD Mahidol and G6PD Kaiping. One subject (0.4%) each carried G6PD Viangchan and G6PD Canton. Interestingly, one case of coinheritance of G6PD Aures and Quing Yan was detected in this cohort. Based on levels of G6PD enzyme activity, the prevalence of G6PD deficiency in the Lao Theung population was 9.52 % (24/252).Conclusion: The G6PD Aures mutation is highly prevalent in the Lao Theung ethnic group. The common G6PD variants in continental Southeast Asian populations, G6PD Viangchan, Canton, Kaiping, Union and Mahidol, were not prevalent in this ethnic group. The technical simplicity of the developed ARMS-PCR will facilitate the final diagnosis of the G6PD Aures.

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