{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,22]],"date-time":"2025-10-22T02:57:36Z","timestamp":1761101856575,"version":"build-2065373602"},"reference-count":24,"publisher":"BMJ","issue":"7","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J Med Genet"],"published-print":{"date-parts":[[1994,7]]},"abstract":"<jats:p>Medium chain acyl-CoA dehydrogenase (MCAD) deficiency is a common inherited metabolic disorder affecting fatty acid beta oxidation. Identification of carriers is important since the disease can be fatal and is readily treatable once diagnosed. Twelve molecular defects have been identified in the MCAD gene; however, a single highly prevalent mutation, A985G, accounts for &gt; 90% of mutant alleles in the white population. In order to facilitate the molecular diagnosis of MCAD deficiency, oligonucleotide primers were designed to amplify the exon regions encompassing the 12 mutations enzymatically, and PCR products were then screened with a single strand conformation polymorphism (SSCP) based method. Minigels were used allowing much faster run times, and silver staining was used after gel electrophoresis to eliminate the need for radioisotopic labelling strategies. Our non-radioactive, minigel SSCP approach showed that normals can be readily distinguished from heterozygotes and homozygotes for all three of the 12 known MCAD mutations which were detected in our sampling of 48 persons. In addition, each band pattern is characteristic for a specific mutation, including those mapping in the same PCR product like A985G and T1124C. When necessary, the molecular defect was confirmed using either restriction enzyme digestion of PCR products or by direct DNA sequence analysis or both. This rapid, non-radioactive approach can become routine for molecular diagnosis of MCAD deficiency and other genetic disorders.<\/jats:p>","DOI":"10.1136\/jmg.31.7.551","type":"journal-article","created":{"date-parts":[[2008,11,10]],"date-time":"2008-11-10T15:23:16Z","timestamp":1226330596000},"page":"551-554","source":"Crossref","is-referenced-by-count":18,"title":["Rapid detection of medium chain acyl-CoA dehydrogenase gene mutations by non-radioactive, single strand conformation polymorphism minigels."],"prefix":"10.1136","volume":"31","author":[{"given":"A","family":"Iolascon","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T","family":"Parrella","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S","family":"Perrotta","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"O","family":"Guardamagna","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"P M","family":"Coates","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M","family":"Sartore","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S","family":"Surrey","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"P","family":"Fortina","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"239","published-online":{"date-parts":[[1994,7,1]]},"reference":[{"key":"ref_1","first-page":"889","article-title":"Acyl CoA dehydrogenase deficiencies","volume-title":"The metabolic basis of inherited diseases","author":"Roe, C.R.; Coates, P.M.","year":"1989","unstructured":"Roe CR, Coates PM. Acyl CoA dehydrogenase deficiencies. In: Scriver CR, Beaudet AL, Sly WS, Valley D, eds. The metabolic basis of inherited diseases. 6th ed. New York: McGraw-Hill, 1989:889-914.","edition":"6th ed"},{"key":"ref_2","first-page":"1280","article-title":"Molecular survey of a prevalent mutation, 985A-to-G transition, and identification offive infrequent mutations in the medium-chain acylCoA dehydrogenase gene in 55 patients with mediumchain acyl-CoA dehydrogenase deficiency","volume":"47","author":"Yokota, Y.; Coates, P.M.; Hale, D.E.","year":"1991","unstructured":"Yokota Y, Coates PM, Hale DE, et al. Molecular survey of a prevalent mutation, 985A-to-G transition, and identification offive infrequent mutations in the medium-chain acylCoA dehydrogenase gene in 55 patients with mediumchain acyl-CoA dehydrogenase deficiency. 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Medium chain acylCoA dehydrogenase deficiency: diagnosis by stable-isotope dilution measurement of urinary n-hexanoylglycine and 3-phenylpropionylglycine. N Engi Med 1988;319: 1308-13.","journal-title":"N Engi Med"},{"key":"ref_12","first-page":"123","article-title":"Inherited disorders of mitochondrial fatty acid oxidation","volume-title":"Progress in liver diseases. Vol X","author":"Coates, P.M.; Stanley, C.A.","year":"1992","unstructured":"Coates PM, Stanley CA. Inherited disorders of mitochondrial fatty acid oxidation. In: Boyer JL, Ockner RK, eds. Progress in liver diseases. Vol X. Philadelphia: Saunders, 1992:123-38."},{"key":"ref_13","first-page":"6677","article-title":"Nucleotide sequence of medium-chain acyl-CoA dehydrogenase mRNA and its expression","volume-title":"enzyme-deficient human tissue. Proc Natl Acad Sci","volume":"86","author":"Kelly, D.P.; Kim, J.J.P.; Billadello, J.J.","year":"1987","unstructured":"Kelly DP, Kim JJP, Billadello JJ, et al. 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Molecular characterization of inherited medium-chain acyl-CoA dehydrogenase deficiency. Proc Natl Acad Sci USA 1990; 87:9236-40.","journal-title":"Proc Natl Acad Sci USA"},{"key":"ref_17","first-page":"54551","article-title":"Molecular characterization of the medium-chain acyl-CoA dehydrogenase (MCAD) deficiency: identification of a Lys\"'9 to Glu mutation in the MCAD gene, and expression of inactive mutant enzyme protein in E. coli","volume":"86","author":"Gregersen, N.; Andresen, B.S.; Bross, P.","year":"1991","unstructured":"Gregersen N, Andresen BS, Bross P, et al. Molecular characterization of the medium-chain acyl-CoA dehydrogenase (MCAD) deficiency: identification of a Lys\"'9 to Glu mutation in the MCAD gene, and expression of inactive mutant enzyme protein in E. coli. Hum Genet 1991;86:54551.","journal-title":"Hum Genet"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"552","DOI":"10.1016\/0140-6736(91)91110-G","article-title":"Prevalence of K329E mutation in medium-chain acyl-CoA dehydrogenase gene determined from Guthrie cards","volume":"338","author":"Matsubara, Y.; Narisawa, K.; Tada, K.","year":"1991","unstructured":"Matsubara Y, Narisawa K, Tada K, et al. Prevalence of K329E mutation in medium-chain acyl-CoA dehydrogenase gene determined from Guthrie cards. Lancet 1991;338:552-4.","journal-title":"Lancet"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1000","DOI":"10.1172\/JCI114761","article-title":"Molecular basis of medium chain acyl-coenzyme A dehydrogenase deficiency. 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Science 1987;239:487-91.","journal-title":"Science"}],"container-title":["Journal of Medical Genetics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1136\/jmg.31.7.551","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,18]],"date-time":"2025-10-18T07:38:08Z","timestamp":1760773088000},"score":1,"resource":{"primary":{"URL":"https:\/\/jmg.bmj.com\/lookup\/doi\/10.1136\/jmg.31.7.551"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1994,7]]},"references-count":24,"journal-issue":{"issue":"7","published-online":{"date-parts":[[1994,7,1]]},"published-print":{"date-parts":[[1994,7]]}},"alternative-id":["10.1136\/jmg.31.7.551"],"URL":"https:\/\/doi.org\/10.1136\/jmg.31.7.551","relation":{},"ISSN":["0022-2593","1468-6244"],"issn-type":[{"type":"print","value":"0022-2593"},{"type":"electronic","value":"1468-6244"}],"subject":[],"published":{"date-parts":[[1994,7]]}}}