Heneine W, Switzer W M
Retrovirus Diseases Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
Transplantation. 1996 Nov 15;62(9):1360-2. doi: 10.1097/00007890-199611150-00033.
Pig and baboon xenotransplants in humans require assays that discriminate source from human cells to investigate engraftment and identify true infection of recipients with xenogeneic endogenous retroviruses. We developed two polymerase chain reaction assays that target a porcine-specific sequence in the beta-globin gene and a baboon-specific sequence in the mitochondrial cytochrome oxidase subunit II gene. The sensitivity and specificity of both assays were evaluated on DNA lysates from baboon, pig, and human peripheral blood lymphocytes. Both assays detected single cells in backgrounds of human DNA. Additionally, both assays were highly specific and yielded negative results in reactions containing only human DNA. The two assays reliably detected peripheral blood lymphocyte samples from 14 baboons and 16 pigs. The baboon- and pig-specific target sequences are gender independent and nonpolymorphic and allow universal applicability. The high sensitivity and specificity is of particular importance in assessing low-level engraftment in human xenotransplant chimeras.
猪和狒狒异种移植到人体需要进行能区分来源细胞与人类细胞的检测，以研究植入情况并确定受者是否真正感染了异种内源性逆转录病毒。我们开发了两种聚合酶链反应检测方法，一种针对β-珠蛋白基因中的猪特异性序列，另一种针对线粒体细胞色素氧化酶亚基II基因中的狒狒特异性序列。这两种检测方法的灵敏度和特异性均在来自狒狒、猪和人类外周血淋巴细胞的DNA裂解物上进行了评估。两种检测方法都能在人类DNA背景中检测到单个细胞。此外，两种检测方法都具有高度特异性，在仅含人类DNA的反应中产生阴性结果。这两种检测方法可靠地检测了来自14只狒狒和16头猪的外周血淋巴细胞样本。狒狒和猪特异性靶序列与性别无关且无多态性，具有普遍适用性。高灵敏度和特异性在评估人类异种移植嵌合体中的低水平植入情况时尤为重要。
