Unbiased targeted next-generation sequencing molecular approach for primary immunodeficiency diseases - 03/06/16
, Mohamed Abouelhoda, PhD b, d, Dorota M. Monies, PhD b, d, Nada Al-Tassan, PhD b, d, Abdulaziz Al-Ghonaium, MD a, Bandar Al-Saud, MD a, c, Hasan Al-Dhekri, MD a, Rand Arnaout, MD a, Saleh Al-Muhsen, MD a, e, Nazema Ades, BN a, Sahar Elshorbagi, MD a, Sulaiman Al Gazlan, MD f, Farrukh Sheikh, MD f, Majed Dasouki, MD b, Lina El-Baik, BSc b, Tanzeil Elamin, MS b, Amal Jaber, BSc b, Omnia Kheir, BPharm b, Mohamed El-Kalioby, MS b, Shazia Subhani, MS b, d, Eman Al Idrissi, MD g, Mofareh Al-Zahrani, MD g, Maryam Alhelale, MD h, Noukha Alnader, BSc b, Afaf Al-Otaibi, BSc b, Rana Kattan, BSc d, Khalid Al Abdelrahman, BSc d, Muna M. Al Breacan, BSc b, Faisal S. Bin Humaid, BSc b, Salma Majid Wakil, PhD b, Fadi Alzayer, BSc i, Haya Al-Dusery, BSc b, Tariq Faquih, MS b, Safa Al-Hissi, BSc b, Brian F. Meyer, PhD b, d, Abbas Hawwari, PhD b, ⁎ Abstract |
Background |
Molecular genetics techniques are an essential diagnostic tool for primary immunodeficiency diseases (PIDs). The use of next-generation sequencing (NGS) provides a comprehensive way of concurrently screening a large number of PID genes. However, its validity and cost-effectiveness require verification.
Objectives |
We sought to identify and overcome complications associated with the use of NGS in a comprehensive gene panel incorporating 162 PID genes. We aimed to ascertain the specificity, sensitivity, and clinical sensitivity of the gene panel and its utility as a diagnostic tool for PIDs.
Methods |
A total of 162 PID genes were screened in 261 patients by using the Ion Torrent Proton NGS sequencing platform. Of the 261 patients, 122 had at least 1 known causal mutation at the onset of the study and were used to assess the specificity and sensitivity of the assay. The remaining samples were from unsolved cases that were biased toward more phenotypically and genotypically complicated cases.
Results |
The assay was able to detect the mutation in 117 (96%) of 122 positive control subjects with known causal mutations. For the unsolved cases, our assay resulted in a molecular genetic diagnosis for 35 of 139 patients. Interestingly, most of these cases represented atypical clinical presentations of known PIDs.
Conclusions |
The targeted NGS PID gene panel is a sensitive and cost-effective diagnostic tool that can be used as a first-line molecular assay in patients with PIDs. The assay is an alternative choice to the complex and costly candidate gene approach, particularly for patients with atypical presentation of known PID genes.
Le texte complet de cet article est disponible en PDF.Key words : Immunodeficiency, next-generation sequencing, targeted, genetic, primary immunodeficiency, mutation, variants, Saudi, diagnosis
Abbreviations used : CID, CNV, ePCR, HGMD, indel, NGS, PID, SCID, SNP, SNV, WES, WGS
Plan
| Supported by the National Science, Technology and Innovation Plan strategic technologies program in Saudi Arabia (KACST: 11-BIO-1441-20). |
|
| Disclosure of potential conflict of interest: H. Al-Mousa, L. El-Baik, O. Khier, A. Al-Otaibi, and S. Al-Hissi have received a grant from King Abdulaziz City for Science and Technology (grant no. KACST: 11-BIO-1441-20). The rest of the authors declare that they have no relevant conflicts of interest. |
Vol 137 - N° 6
P. 1780-1787 - juin 2016 Retour au numéro
Article précédent
- No long-term evidence of hyporesponsiveness after use of pneumococcal conjugate vaccine in children previously immunized with pneumococcal polysaccharide vaccine
- Paul V. Licciardi, Zheng Quan Toh, Elizabeth A. Clutterbuck, Anne Balloch, Rachel A. Marimla, Leena Tikkanen, Karen E. Lamb, Kathryn J. Bright, Uraia Rabuatoka, Lisi Tikoduadua, Laura K. Boelsen, Eileen M. Dunne, Catherine Satzke, Yin Bun Cheung, Andrew J. Pollard, Fiona M. Russell, Edward K. Mulholland
- Changes in IgE sensitization and total IgE levels over 20 years of follow-up
- André F.S. Amaral, Roger B. Newson, Michael J. Abramson, Josep M. Antó, Roberto Bono, Angelo G. Corsico, Roberto de Marco, Pascal Demoly, Bertil Forsberg, Thorarinn Gislason, Joachim Heinrich, Ismael Huerta, Christer Janson, Rain Jõgi, Jeong-Lim Kim, José Maldonado, Jesús Martinez-Moratalla Rovira, Catherine Neukirch, Dennis Nowak, Isabelle Pin, Nicole Probst-Hensch, Chantal Raherison-Semjen, Cecilie Svanes, Isabel Urrutia Landa, Ronald van Ree, Serge A. Versteeg, Joost Weyler, Jan-Paul Zock, Peter G.J. Burney, Deborah L. Jarvis
Bienvenue sur EM-consulte, la référence des professionnels de santé.
L’accès au texte intégral de cet article nécessite un abonnement.
Déjà abonné à cette revue ?