{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T21:24:33Z","timestamp":1791494673799,"version":"4.3.4"},"reference-count":42,"publisher":"BMJ","issue":"2","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J Med Genet"],"accepted":{"date-parts":[[2010,9,13]]},"published-print":{"date-parts":[[2011,2]]},"abstract":"<jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Nephronophthisis associated ciliopathies (NPHP-AC) comprise a group of autosomal recessive cystic kidney diseases that includes nephronophthisis (NPHP), Senior-Loken syndrome (SLS), Joubert syndrome (JBTS), and Meckel-Gruber syndrome (MKS). To date, causative mutations in NPHP-AC have been described for 18 different genes, rendering mutation analysis tedious and expensive. To overcome the broad genetic locus heterogeneity, a strategy of DNA pooling with consecutive massively parallel resequencing (MPR) was devised.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>In 120 patients with severe NPHP-AC phenotypes, five pools of genomic DNA with 24 patients each were prepared which were used as templates in order to PCR amplify all 376 exons of 18 NPHP-AC genes (<jats:italic>NPHP1<\/jats:italic>, <jats:italic>INVS<\/jats:italic>, <jats:italic>NPHP3<\/jats:italic>, <jats:italic>NPHP4<\/jats:italic>, <jats:italic>IQCB1<\/jats:italic>, <jats:italic>CEP290<\/jats:italic>, <jats:italic>GLIS2<\/jats:italic>, <jats:italic>RPGRIP1L<\/jats:italic>, <jats:italic>NEK8<\/jats:italic>, <jats:italic>TMEM67<\/jats:italic>, <jats:italic>INPP5E<\/jats:italic>, <jats:italic>TMEM216<\/jats:italic>, <jats:italic>AHI1<\/jats:italic>, <jats:italic>ARL13B<\/jats:italic>, <jats:italic>CC2D2A<\/jats:italic>, <jats:italic>TTC21B<\/jats:italic>, <jats:italic>MKS1<\/jats:italic>, and <jats:italic>XPNPEP3<\/jats:italic>). PCR products were then subjected to MPR on an Illumina Genome-Analyser and mutations were subsequently assigned to their respective mutation carrier via CEL I endonuclease based heteroduplex screening and confirmed by Sanger sequencing.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>For proof of principle, DNA from patients with known mutations was used and detection of 22 out of 24 different alleles (92% sensitivity) was demonstrated. MPR led to the molecular diagnosis in 30\/120 patients (25%) and 54 pathogenic mutations (27 novel) were identified in seven different NPHP-AC genes. Additionally, in 24 patients only single heterozygous variants of unknown significance were found.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>The combined approach of DNA pooling followed by MPR strongly facilitates mutation analysis in broadly heterogeneous single gene disorders. The lack of mutations in 75% of patients in this cohort indicates further extensive heterogeneity in NPHP-AC.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1136\/jmg.2010.082552","type":"journal-article","created":{"date-parts":[[2010,11,10]],"date-time":"2010-11-10T21:01:42Z","timestamp":1289422902000},"page":"105-116","source":"Crossref","is-referenced-by-count":123,"title":["Mutation analysis of 18 nephronophthisis associated ciliopathy disease genes using a DNA pooling and next generation sequencing strategy"],"prefix":"10.1136","volume":"48","author":[{"given":"Edgar 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