An effective combination of sanger and next generation sequencing in diagnostics of primary ciliary dyskinesia
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00064203%3A_____%2F16%3A10323843" target="_blank" >RIV/00064203:_____/16:10323843 - isvavai.cz</a>
Alternative codes found
RIV/00216208:11130/16:10323843
Result on the web
<a href="http://dx.doi.org/10.1002/ppul.23261" target="_blank" >http://dx.doi.org/10.1002/ppul.23261</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1002/ppul.23261" target="_blank" >10.1002/ppul.23261</a>
Alternative languages
Result language
angličtina
Original language name
An effective combination of sanger and next generation sequencing in diagnostics of primary ciliary dyskinesia
Original language description
BackgroundPrimary ciliary dyskinesia (PCD) is a multigenic autosomal recessive condition affecting respiratory tract and other organs where ciliary motility is required. The extent of its genetic heterogeneity is remarkable. The aim of the study was to develop a cost-effective pipeline for genetic diagnostics using a combination of Sanger and next generation sequencing (NGS). Materials and MethodsData and samples of 33 families with 38 affected subjects with PCD diagnosed in childhood were collected over the territory of the Czech Republic. A panel of 18 PCD causative or candidate genes was implemented into an Illumina TruSeq Custom Amplicon NGS assay, and three ancestral mutations in SPAG1 were screened by conventional Sanger sequencing, which was also used for the confirmation of the NGS results and for the analysis of familial segregation. ResultsThe causative gene was DNAH5 in 11/33 (33%) probands, SPAG1 in 8/33 (24%), and DNAI1, CCDC40, LRRC6 in one family each. If the high proportion of subjects with bi-allelic ancestral mutations in SPAG1 is corroborated in other Caucasian populations, a simple Sanger sequencing test for these three mutations may serve as an effective pre-screening step, being followed by an NGS panel for other, much larger, PCD genes. ConclusionsWe present a combination of Sanger sequencing with an NGS panel for known and candidate PCD genes, implemented in a moderate-size national collection of patients. This strategy has proven to be cost-effective, rapid and reliable, and was able to detect the causative gene in two thirds of our PCD patients. Pediatr Pulmonol. 2016;51:498-509. (c) 2015 Wiley Periodicals, Inc.
Czech name
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Czech description
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Classification
Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
FG - Paediatrics
OECD FORD branch
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Result continuities
Project
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Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2016
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Pediatric Pulmonology
ISSN
8755-6863
e-ISSN
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Volume of the periodical
51
Issue of the periodical within the volume
5
Country of publishing house
US - UNITED STATES
Number of pages
12
Pages from-to
498-509
UT code for WoS article
000374303600009
EID of the result in the Scopus database
2-s2.0-84963837836