Roth J, Daus H, Gause A, Trümper L, Pfreundschuh M
Innere Medizin I, Universitätskliniken des Saarlandes, Homburg, Germany.
Leuk Lymphoma. 1994 Mar;13(1-2):137-42. doi: 10.3109/10428199409051664.
The Epstein-Barr virus (EBV) can be detected in the majority of lymph nodes involved by Hodgkin's lymphoma using the highly sensitive polymerase chain reaction (PCR). However, the rate of EBV-DNA detection by in-situ hybridisation, which allows allocation of EBV to a defined cell population, i.e. the neoplastic H&RS-cells, is lower. In an attempt to combine the advantages of the high sensitivity of the PCR and the possibility of cellular allocation by in-situ hybridisation, we established a single-cell PCR of Hodgkin- and Reed-Sternberg (H&RS)-cells isolated by micromanipulation from biopsy tissues. We amplified EBV sequences from the BamW-region by single-cell PCR. Using this method we were able to detect EBV-DNA in the H&RS-cells from 4 of 6 patients. In EBV positive cases all H&RS-cells of a given patient were positive, proving the high sensitivity and reproducibility of the method. Other cells in the biopsy tissue involved by EBV-positive H&RS-cells were shown to be negative. This indicates that EBV may have a role in the pathogenesis of many but not all cases of Hodgkin's disease.
利用高灵敏度的聚合酶链反应（PCR），可在大多数霍奇金淋巴瘤累及的淋巴结中检测到爱泼斯坦-巴尔病毒（EBV）。然而，原位杂交检测EBV-DNA的比率较低，原位杂交可将EBV定位到特定细胞群体，即肿瘤性霍奇金和里德-斯腾伯格（H&RS）细胞。为了结合PCR高灵敏度的优势以及原位杂交进行细胞定位的可能性，我们建立了一种从活检组织中通过显微操作分离出的霍奇金和里德-斯腾伯格（H&RS）细胞的单细胞PCR方法。我们通过单细胞PCR扩增了BamW区域的EBV序列。使用该方法，我们能够在6例患者中的4例的H&RS细胞中检测到EBV-DNA。在EBV阳性病例中，给定患者的所有H&RS细胞均为阳性，证明了该方法的高灵敏度和可重复性。活检组织中被EBV阳性H&RS细胞累及的其他细胞显示为阴性。这表明EBV可能在许多但并非所有霍奇金病病例的发病机制中起作用。
