{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T15:13:32Z","timestamp":1764688412813},"reference-count":27,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2004,10,13]],"date-time":"2004-10-13T00:00:00Z","timestamp":1097625600000},"content-version":"vor","delay-in-days":3239,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Muscle and Nerve"],"published-print":{"date-parts":[[1995,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>We have devised a novel method for quantitative analysis of the MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis, and strokelike episodes) tRNA<jats:sup>Leu(UUR)<\/jats:sup> mutation of mitochondrial DNA using a PCR\u2010SSCP (polymerase chain reaction\u2013single\u2010strand conformation polymorphism) method, and compared the results obtained using the PCR\u2010SSCP method with those obtained using other methods including Southern blotting, last one cycle hot PCR, and conventional PCR\u2010RFLP (restriction fragment length polymorphism). The standard curve obtained using the PCR\u2010SSCP method is linear, with a correlation coefficient of 0.999; it was determined that this method is more accurate than other methods for quantitative analysis. The PCR\u2010SSCP method does not require restriction digestions, thereby avoiding potential problems of partial digestions or heteroduplex formation during PCR. The method is quite simple and should have a broad range of application for quantitation of mutant mtDNAs in various mitochondrial encephalo\u2010myopathies. We applied the method for quantitation of mutant mitochondrial DNA carrying a single base substitution in the tRNA<jats:sup>Leu(UUR)<\/jats:sup> gene in two autopsied cases of MELAS. In both cases, the mutant mtDNA is abundantly present (82\u201395%) with little variation among tissues. \u00a9 1995 John Wiley &amp; Sons, Inc.<\/jats:p>","DOI":"10.1002\/mus.880181208","type":"journal-article","created":{"date-parts":[[2005,1,1]],"date-time":"2005-01-01T02:32:15Z","timestamp":1104546735000},"page":"1390-1397","source":"Crossref","is-referenced-by-count":14,"title":["Quantitation of heteroplasmy of mitochondrial trna<sup>Leu(UUR)<\/sup> gene using PCR\u2010SSCP"],"prefix":"10.1002","volume":"18","author":[{"given":"Yoshinori","family":"Tanno","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Makoto","family":"Yoneda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keiko","family":"Tanaka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hajime","family":"Tanaka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Motoyoshi","family":"Yamazaki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masatoyo","family":"Nishiawa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Koichi","family":"Wakabayashi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eisaku","family":"Ohama","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shoji","family":"Tsuji","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2004,10,13]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/290457a0"},{"key":"e_1_2_1_3_2","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/S0021-9258(19)36552-4","article-title":"Abnormal RNA processing associated with a novel tRNA mutation in mitochondrial DNA. A potential disease mechanism","volume":"268","author":"Bindoff LA","year":"1993","journal-title":"J Biol Chem"},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.89.10.4221"},{"key":"e_1_2_1_5_2","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.41.10.1663"},{"key":"e_1_2_1_6_2","doi-asserted-by":"publisher","DOI":"10.1002\/ana.410310408"},{"key":"e_1_2_1_7_2","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.42.3.545"},{"key":"e_1_2_1_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/348651a0"},{"key":"e_1_2_1_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/331717a0"},{"key":"e_1_2_1_10_2","first-page":"428","article-title":"A new mitochondrial disease associated with mitochondrial DNA heteroplasmy","volume":"46","author":"Holt IJ","year":"1990","journal-title":"Am J Hum Genet"},{"key":"e_1_2_1_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.86.23.9253"},{"key":"e_1_2_1_12_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01800025"},{"key":"e_1_2_1_13_2","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.43.8.1586"},{"key":"e_1_2_1_14_2","first-page":"934","article-title":"The mitochondrial tRNALeu(UUR) mutation in mitochondrial encephalomyopathy, lactic acidosis, and strokelike episodes (MELAS): genetic, biochemical, and morphological correlations in skeletal muscle","volume":"50","author":"Moraes CT","year":"1992","journal-title":"Am J Hum Genet"},{"key":"e_1_2_1_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/ng0892-359"},{"key":"e_1_2_1_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(91)80942-V"},{"key":"e_1_2_1_17_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF00688185"},{"key":"e_1_2_1_18_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.86.8.2766"},{"key":"e_1_2_1_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291X(88)90272-0"},{"key":"e_1_2_1_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-510X(90)90133-8"},{"key":"e_1_2_1_21_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.2448875"},{"key":"e_1_2_1_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-510X(93)90270-9"},{"key":"e_1_2_1_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(90)90059-N"},{"key":"e_1_2_1_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291X(91)91900-W"},{"key":"e_1_2_1_25_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.3201231"},{"key":"e_1_2_1_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.14.4.2699"},{"key":"e_1_2_1_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-510X(89)90132-9"},{"key":"e_1_2_1_28_2","first-page":"789","article-title":"A common mitochondrial DNA mutation in the t\u2010RNALys of patients with myoclonus epilepsy associated with ragged\u2010red fibers","volume":"21","author":"Yoneda M","year":"1990","journal-title":"Biochem Int"}],"container-title":["Muscle &amp; Nerve"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fmus.880181208","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/mus.880181208","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,26]],"date-time":"2023-10-26T03:12:50Z","timestamp":1698289970000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/mus.880181208"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1995,12]]},"references-count":27,"journal-issue":{"issue":"12","published-print":{"date-parts":[[1995,12]]}},"alternative-id":["10.1002\/mus.880181208"],"URL":"https:\/\/doi.org\/10.1002\/mus.880181208","archive":["Portico"],"relation":{},"ISSN":["0148-639X","1097-4598"],"issn-type":[{"value":"0148-639X","type":"print"},{"value":"1097-4598","type":"electronic"}],"subject":[],"published":{"date-parts":[[1995,12]]}}}