A Simple, Cost-Effective, and Extraction-Free Molecular Diagnostic Test for Sickle Cell Disease Using a Noninvasive Buccal Swab Specimen for a Limited-Resource Setting
Sickle cell disease (SCD) is the most prevalent life-threatening blood monogenic disorder. Currently, there is no cure available, apart from bone marrow transplantation. Early and efficient diagnosis of SCD is key to disease management, which would make considerable strides in alleviating morbidity...
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| Format: | Article |
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MDPI AG
2022-07-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/12/7/1765 |
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| author | Priya Thakur Pragya Gupta Nupur Bhargava Rajat Soni Narendra Varma Gottumukkala Sangam Giri Goswami Gaurav Kharya Vinodh Saravanakumar Padma Gunda Suman Jain Jasmita Dass Mukul Aggarwal Sivaprakash Ramalingam |
| author_facet | Priya Thakur Pragya Gupta Nupur Bhargava Rajat Soni Narendra Varma Gottumukkala Sangam Giri Goswami Gaurav Kharya Vinodh Saravanakumar Padma Gunda Suman Jain Jasmita Dass Mukul Aggarwal Sivaprakash Ramalingam |
| author_sort | Priya Thakur |
| collection | DOAJ |
| description | Sickle cell disease (SCD) is the most prevalent life-threatening blood monogenic disorder. Currently, there is no cure available, apart from bone marrow transplantation. Early and efficient diagnosis of SCD is key to disease management, which would make considerable strides in alleviating morbidity and reducing mortality. However, the cost and complexity of diagnostic procedures, such as the Sanger sequencing method, impede the early detection of SCD in a resource-limited setting. To address this, the current study demonstrates a simple and efficient proof-of-concept assay for the detection of patients and carriers using extraction-free non-invasive buccal swab samples by isothermal DNA Amplification coupled Restrictase-mediated cleavage (iDAR). This study is a first of its kind reporting the use of buccal swab specimens for iDA in molecular diagnosis of a genetic disease, all the while being cost effective and time saving, with the total assay time of around 150 min at a cost of USD 5. Further, iDAR demonstrates 91.5% sensitivity and 100% specificity for detecting all three alleles: SS, AS, and AA, having a 100% concordance with Sanger sequencing. The applicability of the iDAR assay is further demonstrated with its adaptation to a one-pot reaction format, which simplifies the assay system. Overall, iDAR is a simple, cost-effective, precise, and non-invasive assay for SCD screening, with the potential for use in a limited resource setting. |
| first_indexed | 2024-03-09T10:20:12Z |
| format | Article |
| id | doaj.art-53809a58ee74487d832c6189943ff76d |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-03-09T10:20:12Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-53809a58ee74487d832c6189943ff76d2023-12-01T22:04:27ZengMDPI AGDiagnostics2075-44182022-07-01127176510.3390/diagnostics12071765A Simple, Cost-Effective, and Extraction-Free Molecular Diagnostic Test for Sickle Cell Disease Using a Noninvasive Buccal Swab Specimen for a Limited-Resource SettingPriya Thakur0Pragya Gupta1Nupur Bhargava2Rajat Soni3Narendra Varma Gottumukkala4Sangam Giri Goswami5Gaurav Kharya6Vinodh Saravanakumar7Padma Gunda8Suman Jain9Jasmita Dass10Mukul Aggarwal11Sivaprakash Ramalingam12CSIR—Institute Genomics and Integrative Biology, Mathura Road, Sukhdev Vihar, New Delhi 110025, IndiaCSIR—Institute Genomics and Integrative Biology, Mathura Road, Sukhdev Vihar, New Delhi 110025, IndiaCSIR—Institute Genomics and Integrative Biology, Mathura Road, Sukhdev Vihar, New Delhi 110025, IndiaCSIR—Institute Genomics and Integrative Biology, Mathura Road, Sukhdev Vihar, New Delhi 110025, IndiaCSIR—Institute Genomics and Integrative Biology, Mathura Road, Sukhdev Vihar, New Delhi 110025, IndiaCSIR—Institute Genomics and Integrative Biology, Mathura Road, Sukhdev Vihar, New Delhi 110025, IndiaIndraprastha Apollo Hospitals, Sarita Vihar, New Delhi 110020, IndiaCSIR—Institute Genomics and Integrative Biology, Mathura Road, Sukhdev Vihar, New Delhi 110025, IndiaThalassemia and Sickle Cell Society, Rajendra Nagar, Hyderabad 500052, IndiaThalassemia and Sickle Cell Society, Rajendra Nagar, Hyderabad 500052, IndiaDepartment of Hematology, All India Institute of Medical Sciences, New Delhi 110029, IndiaDepartment of Hematology, All India Institute of Medical Sciences, New Delhi 110029, IndiaCSIR—Institute Genomics and Integrative Biology, Mathura Road, Sukhdev Vihar, New Delhi 110025, IndiaSickle cell disease (SCD) is the most prevalent life-threatening blood monogenic disorder. Currently, there is no cure available, apart from bone marrow transplantation. Early and efficient diagnosis of SCD is key to disease management, which would make considerable strides in alleviating morbidity and reducing mortality. However, the cost and complexity of diagnostic procedures, such as the Sanger sequencing method, impede the early detection of SCD in a resource-limited setting. To address this, the current study demonstrates a simple and efficient proof-of-concept assay for the detection of patients and carriers using extraction-free non-invasive buccal swab samples by isothermal DNA Amplification coupled Restrictase-mediated cleavage (iDAR). This study is a first of its kind reporting the use of buccal swab specimens for iDA in molecular diagnosis of a genetic disease, all the while being cost effective and time saving, with the total assay time of around 150 min at a cost of USD 5. Further, iDAR demonstrates 91.5% sensitivity and 100% specificity for detecting all three alleles: SS, AS, and AA, having a 100% concordance with Sanger sequencing. The applicability of the iDAR assay is further demonstrated with its adaptation to a one-pot reaction format, which simplifies the assay system. Overall, iDAR is a simple, cost-effective, precise, and non-invasive assay for SCD screening, with the potential for use in a limited resource setting.https://www.mdpi.com/2075-4418/12/7/1765molecular diagnostic testsickle cell diseasenon-invasiveisothermal DNA amplificationbuccal swabextraction-free |
| spellingShingle | Priya Thakur Pragya Gupta Nupur Bhargava Rajat Soni Narendra Varma Gottumukkala Sangam Giri Goswami Gaurav Kharya Vinodh Saravanakumar Padma Gunda Suman Jain Jasmita Dass Mukul Aggarwal Sivaprakash Ramalingam A Simple, Cost-Effective, and Extraction-Free Molecular Diagnostic Test for Sickle Cell Disease Using a Noninvasive Buccal Swab Specimen for a Limited-Resource Setting Diagnostics molecular diagnostic test sickle cell disease non-invasive isothermal DNA amplification buccal swab extraction-free |
| title | A Simple, Cost-Effective, and Extraction-Free Molecular Diagnostic Test for Sickle Cell Disease Using a Noninvasive Buccal Swab Specimen for a Limited-Resource Setting |
| title_full | A Simple, Cost-Effective, and Extraction-Free Molecular Diagnostic Test for Sickle Cell Disease Using a Noninvasive Buccal Swab Specimen for a Limited-Resource Setting |
| title_fullStr | A Simple, Cost-Effective, and Extraction-Free Molecular Diagnostic Test for Sickle Cell Disease Using a Noninvasive Buccal Swab Specimen for a Limited-Resource Setting |
| title_full_unstemmed | A Simple, Cost-Effective, and Extraction-Free Molecular Diagnostic Test for Sickle Cell Disease Using a Noninvasive Buccal Swab Specimen for a Limited-Resource Setting |
| title_short | A Simple, Cost-Effective, and Extraction-Free Molecular Diagnostic Test for Sickle Cell Disease Using a Noninvasive Buccal Swab Specimen for a Limited-Resource Setting |
| title_sort | simple cost effective and extraction free molecular diagnostic test for sickle cell disease using a noninvasive buccal swab specimen for a limited resource setting |
| topic | molecular diagnostic test sickle cell disease non-invasive isothermal DNA amplification buccal swab extraction-free |
| url | https://www.mdpi.com/2075-4418/12/7/1765 |
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