Testing for Familial Pancreatitis家族性胰腺炎的诊断测试课件.ppt

Diagnostic Testing for Familial Pancreatitis Emma McCarthy Clinical Scientist Merseyside Cheshire Regional Molecular Genetics Laboratory Aims Overview of pancreatitis both hereditary and idiopathic Genes involved diagnostic service PRSS1 dosage analysis Pancreatitis Inflammatory disease Severe abdominal pain Nausea Vomiting fever Acute or chronic Premature activation digestive enzymes Common triggers Excessive alcohol consumption Smoking Gallstones TRYPSINOGEN TRYPSIN AUTODIGESTION CHRONIC PANCREATITIS PERMANENT LOSS OF FUNCTION DIABETES / PANCREATIC CANCER Diagnostic service PRSS1 Fluorescent sequencing exons 2 + 3 SPINK1 p.N34S mutation (pyrosequencing) CFTR 28 mutations (Elucigene CFHT) EUROPAC: www.liv.ac.uk/surgery/europac.html Hereditary Pancreatitis (HP) Autosomal dominant Reduced penetrance Childhood: recurrent episodes acute pancreatitis Adulthood: chronic pancreatitis Mutations of cationic trypsinogen gene (PRSS1) Cationic Trypsinogen PRSS1 gene (protease serine 1) Trypsin Activates digestive enzymes Self regulating Autocatalytic activation Feedback inhibition - cleaves at Arg122 R122H mutation - “super trypsin” Laboratory Data Prior 2007: PCR / restriction digest - p.A16V, p.N29I, p.R122H Feb 2007: fluorescent sequencing exons 2 3 Pathogenic mutations detected - 7/77 (9.1%) 2/77 unclassified variants Total sequence changes identified - 9/77 (11.7%) Idiopathic Chronic Pancreatitis (ICP) 30% pancreatitis cases - unknown cause Susceptibility genes: SPINK1 - Serine Protease Inhibitor Kazal type 1 CFTR – mild mutations; ICP specific mutations? ?polygenic model SPINK1 Inhibits 20% pancreatic trypsin p.N34S mutation 22% ICP patients 2.5% general population ?disease modifier SPINK1 Analysis Pyrosequencing assay c.101AG; p.N34S 9/60 mutation carriers 0/8 CFTR mutations no CF testing on 1 positive case PRSS1 Dosage Analysis Triplication of trypsinogen locus reported Le Maréchal et al., Nature Genetics 2006; 38:1372-4 Fluore