一种新颖的抗原信号增强方法─BT法在免疫组织化学中的应用

本文报道了一种新颖的抗原信号增强方法在免疫组织化学中的应用。该方法的特点是利用生物素标记的酪氨（ＢＴ）结合到ＨＲＰ的催化位点上来达到增强信号的目的。本研究以冰冻切片及石蜡切片中ＡＢＣ法为对照,结合微波和蛋白酶消化预处理,采用ＢＴ法检测了人扁桃体石蜡切片标本中淋巴细胞ＩｇＤ的表达。结果表明该方法检测敏感性高,当一抗稀释至１：５０００时仍能测得ＩｇＤ的表达,与对照的ＡＢＣ法相比,检测敏感性成百倍地提高。该方法能将原先只能用于冰冻切片的单抗ＩｇＤ应用于石蜡切片中。本文还对该方法的可能机制进行了探讨。本研究提示ＢＴ法在病理检验和医学研究中有良好的应用前景。

A NOVEL IMMUNOHISTOCHEMICAL METHOD-BT METHOD FOR ENHANCEMENT OF ANTIGEN SIGNAL I. Detection of IgD expression in lyhiphocyte on paraffin-embedded section with BT method

Here we report the application of a novel signal enhancement method in immunohistochemistry. A key feature of BT method is that biotin-labeling tyramine (BT) is appliedand connected to the sites catalyzed by HRP which has been hanged on the second antibody,so that antigen signal recognized by primary antibody is enhanced. Cryostat and paraffin section are stained with ABC technique in comparison with the BT staining method. lgD expression in lymphocytes on paraffin section of human tonsils is detected with BT method combined with the pretreatments of pronase E digestion, microwave radiation and some detergents. The result shows that BT method is a powerful immunohistochemical staining protocol with high sensitivity. IgD expression in lymphocytes can be detected with better discernment when the dilution of the primary antibody reachs to 1: 5000. There is more than ahundred times enhancement of sensitivity for BT method in comparison with the ABC technique. Our research also presents that the method makes it possible that monoclonal antibody IgD originally used for cryostat section can be used for paraffin section. Finally, thepossible mechanism of BT method for enhancement of antigens signals is discussed. Our research suggests that the practical application of BT method may open up"'broad prospects forthe pathological examination and medical research.