人类中期染色体三色引物原位标记技术研究

Triple-color primed in situ labeling protocol of human metaphase chromosomes

目的探索应用优化的三色引物原位标记技术同时标记多条人类外周血淋巴细胞培养中期染色体的可行性。方法采用双色引物原位标记预实验检测外周血中期淋巴细胞的X、Y染色体以摸索和优化引物应用条件和反应顺序。以优化的三色技术同时标记正常样本及克氏综合征样本（47，XXY）的18、X、Y染色体以验证其有效性。结果在3h之内特异性的同时标记了18、X、Y染色体，双色和三色反应的标记率均达到90％以上。克氏综合征样本准确显示了2条18号染色体，2条X染色体和1条Y染色体。结论非ddNTP阻断的多色引物原位标记技术能够快速而准确地判别染色体数目异常的外周血淋巴细胞样本，原位标记由于有内对照而更加具有说服力。

Objective To study the feasibility of simultaneous detection for several chromosomes with optimized triple-color primed in situ labelling （PRINS） protocol in cultured peripheral blood lymphocytes. Methods Pre-test of gonosome detection with dual-color PRINS protocol was performed to explore and optimize the order and condition of PRINS primers. A peripheral blood sample from a Klinefelter＇ s syndrome patient （47, XXY） had also been studied with optimized triple-eolor PRINS to prove the correspondence between the number of signals and chromosomes. Results Chromosome 18, X and Y had been simultaneously and specifically marked within 3 hours. The frequency-of successful labeling reached 90% both in dual-color and triple-eolor test. Two chromosome X had been correctly showed in lymphocyte sample of Klinerfelter＇s syndrome. Conclusion Numerical chromosome anomalies could be rapidly and exactly detected with this non-ddNTP-blocking multieolor PRINS protocol in peripheral blood lymphoeytes. The results of in situ labeling are much dearer with inner control.