阅读说明：本技术 通过管内杂交与通用标签-微阵列的组合的突变的基因分型 (Genotyping of mutations by in-tube hybridization in combination with universal tag-microarray ) 是由 玛赛拉·基亚里 弗朗西斯科·达明 西尔维亚·加尔比亚蒂 莫里齐奥·费拉里 于 2018-03-30 设计创作，主要内容包括：目标基因片段的测定方法,其包括：扩增至少一种包含或可能包含至少一种目标SNP区域的目标基因片段,以形成初始扩增产物；从初始扩增产物形成至少一种单链扩增产物,其中所述单链扩增产物包含或可能包含至少一种目标SNP区域；在溶液中将所述单链扩增产物与至少一种报道分子杂交,所述报道分子包含寡核苷酸的至少两个不同的结构域,其中寡核苷酸的第一结构域用于与单链扩增产物杂交,并且其中寡核苷酸的第二结构域用于与至少一种微阵列探针表面杂交,其中微阵列探针表面包含至少一种捕获探针以允许杂交；将杂交单链扩增产物的溶液与至少一种微阵列探针表面接触,所述微阵列探针表面包含至少一种捕获探针以允许杂交；检测微阵列表面上杂交单链扩增产物的存在。在与癌症基因分型相关的优选实施方案中,KRAS癌基因的出人意料的良好结果表明所研究的所有7种密码子12和13突变可在小于90分钟内在组织临床样品中明确检测到。此外,该系统可以揭示代表少于0.1％起始材料的突变等位基因。通过将突变检测与阵列杂交解偶联,该技术变得通用。(A method for assaying a target gene fragment, comprising: amplifying at least one target gene fragment comprising or potentially comprising at least one SNP region of interest to form an initial amplification product; forming at least one single-stranded amplification product from the initial amplification product, wherein the single-stranded amplification product comprises or may comprise at least one SNP region of interest; hybridizing the single-stranded amplification product in solution with at least one reporter molecule comprising at least two different domains of an oligonucleotide, wherein a first domain of an oligonucleotide is used for hybridization with the single-stranded amplification product, and wherein a second domain of an oligonucleotide is used for hybridization with at least one microarray probe surface, wherein the microarray probe surface comprises at least one capture probe to allow hybridization; contacting a solution of hybridized single-stranded amplification products with at least one microarray probe surface comprising at least one capture probe to allow hybridization; detecting the presence of the hybridized single stranded amplification product on the surface of the microarray. In a preferred embodiment associated with cancer genotyping, unexpectedly good results for the KRAS oncogene indicate that all 7 codon 12 and 13 mutations studied can be unambiguously detected in clinical samples of tissue in less than 90 minutes. In addition, the system can reveal mutant alleles that represent less than 0.1% of the starting material. This technique becomes versatile by uncoupling the mutation detection from the array hybridization.)