Musholt P B, Musholt T J, Goodfellow P J, Zehnbauer B A, Wells S A, Moley J F
Department of Surgery, Washington University School of Medicine, St. Louis, Mo. 63110, USA.
Surgery. 1997 Aug;122(2):363-70; discussion 370-1. doi: 10.1016/s0039-6060(97)90028-3.
RET protooncogene mutation analysis is a routinely performed predictive DNA test in kindreds affected by multiple endocrine neoplasia (MEN) types 2A and 2B and familial medullary thyroid carcinoma (FMTC), and is a valuable diagnostic tool in newly diagnosed cases of medullary thyroid carcinoma (MTC).
We tested the suitability of the recently introduced "cold" single-strand conformational variant (SSCV) technique, which promises rapid, simple, nonradioactive detection of sequence variants in the identification of germline and somatic RET mutations. A total of 11 different mutations in exon 10 (codons 609, 611, 618, and 620) and 6 mutations in exon 11 (codon 634) were studied.
Conditions were optimized so that conformational variants were demonstrated for all mutations examined in a single setting for exons 10 and 11. A novel six base pair (bp) inframe deletion between cysteines 630 and 634 was detected in a sporadic MTC. This adds to the evidence that not only cysteine deletions and substitutions but also changes in the spacing between cysteine residues have a pathogenic effect.
Our results indicate that the cold SSCV method offers the advantages of simplicity, time savings, and nonradioactive detection for screening for RET sequence variants in hereditary and sporadic MTCs.
RET原癌基因突变分析是在受2A和2B型多发性内分泌肿瘤（MEN）以及家族性甲状腺髓样癌（FMTC）影响的家族中常规进行的预测性DNA检测，并且是甲状腺髓样癌（MTC）新诊断病例中的一种有价值的诊断工具。
我们测试了最近引入的“冷”单链构象变异（SSCV）技术的适用性，该技术有望在识别种系和体细胞RET突变时快速、简单、非放射性地检测序列变异。研究了外显子10（密码子609、611、618和620）中的11种不同突变以及外显子11（密码子634）中的6种突变。
条件得到优化，以便在单一设置中针对外显子10和11中检测的所有突变展示构象变异。在一例散发性MTC中检测到半胱氨酸630和634之间一个新的6个碱基对（bp）的框内缺失。这进一步证明，不仅半胱氨酸缺失和取代，而且半胱氨酸残基之间间距的改变也具有致病作用。
我们的结果表明，冷SSCV方法在筛查遗传性和散发性MTC中的RET序列变异方面具有简单、省时和非放射性检测的优点。
