O'Briain D S, Sheils O, McElwaine S, McCann S R, Lawler M
Department of Histopathology, St. James's Hospital, Dublin, Ireland.
Am J Clin Pathol. 1996 Dec;106(6):758-64. doi: 10.1093/ajcp/106.6.758.
Standard identification systems usually ensure that biopsy material is correctly associated with a given patient. Sometimes, as when a tumor is unexpectedly found, the provenance (proof of origin) of a tissue sample may be questioned; the tissue may have been mislabelled or contaminated with tissue from another patient. Techniques used to confirm tissue provenance include comparing either tissue markers of gender or ABO blood groups; however, these methods have weak confirmatory power. Recently, the use of DNA-based polymerase chain reaction (PCR) techniques has been reported. Paired, formalin-fixed, paraffin-embedded, 10 microns tissue sections were selected from 17 patients, 8 of whom had carcinoma, either by dividing a biopsy section, using sequential biopsies, or sequential biopsy and autopsy tissue. The resulting 36 samples were coded before analysis. In two additional cases, 1-mm fragments of tumor from one patient were included in the tissue block of benign tissue from another patient, the tumor fragments were identified on hematoxylin-and-eosin-stained sections, separately scraped off the glass slide, and analyzed. Tissue from two clinical cases, one of suspected mislabelling and one with a suspected carry-over of malignant tissue were also investigated. Short tandem repeat sequences (STR) or microsatellites, are 2-5 base pair repeats that vary in their repeat number between individuals. This variation (polymorphism) can be assessed using a PCR. A panel of markers of 3 STRs; ACPP, INT 2, and CYP 19 (on chromosomes 3, 11, and 15, respectively) were used. DNA was isolated from the samples after xylene deparaffinization and proteinase digestion, and was then amplified in a radioactive PCR using primers selected to give a product size ranging from 136-178 bases. Amplified products were electrophoresed on denaturing polyacrylamide gels, dried, and autoradiographed. DNA segments were successfully extracted from all samples but one, which was fixed in Bouin's fluid. By comparing allele sizes from the panel, all tissue pairs (other than the Bouin's pair) were successfully matched, the 1-mm tumor fragments were correctly assigned, and the two clinical problems were solved. STRs are highly informative and robust markers, well suited to PCR of small portions of tissue sections, and are an effective method to confirm the provenance of benign and malignant biopsy and autopsy material.
标准识别系统通常能确保活检材料与特定患者正确关联。有时，比如意外发现肿瘤时，组织样本的来源（起源证明）可能会受到质疑；组织可能被贴错标签或被另一名患者的组织污染。用于确认组织来源的技术包括比较性别组织标志物或ABO血型；然而，这些方法的确认能力较弱。最近，有报道使用基于DNA的聚合酶链反应（PCR）技术。从17名患者中选取成对的、经福尔马林固定、石蜡包埋的10微米组织切片，其中8名患者患有癌症，切片获取方式包括分割活检切片、连续活检或连续活检及尸检组织。所得的36个样本在分析前进行编码。在另外两个病例中，将一名患者的1毫米肿瘤碎片包含在另一名患者的良性组织组织块中，在苏木精 - 伊红染色切片上识别肿瘤碎片，从载玻片上分别刮下并进行分析。还对来自两个临床病例的组织进行了研究，一个怀疑贴错标签，另一个怀疑有恶性组织残留。短串联重复序列（STR）或微卫星是2 - 5个碱基对的重复序列，其重复次数在个体之间有所不同。这种变异（多态性）可以通过PCR进行评估。使用了一组由3个STR组成的标志物；ACPP、INT 2和CYP 19（分别位于染色体3、11和15上）。在二甲苯脱蜡和蛋白酶消化后从样本中分离DNA，然后在放射性PCR中使用选定的引物进行扩增，以产生大小在136 - 178个碱基之间的产物。扩增产物在变性聚丙烯酰胺凝胶上进行电泳、干燥并进行放射自显影。除了一个用Bouin氏液固定的样本外，所有样本均成功提取出DNA片段。通过比较该组标志物的等位基因大小，所有组织对（除Bouin氏液固定的那对组织外）均成功匹配，1毫米肿瘤碎片被正确识别，两个临床问题也得到了解决。STR是信息丰富且稳定的标志物，非常适合对组织切片的小部分进行PCR，是确认良性和恶性活检及尸检材料来源的有效方法。
