Saito I, Servenius B, Compton T, Fox R I
Department of Basic and Clinical Research, Scripps Clinic, La Jolla, California 92037.
J Exp Med. 1989 Jun 1;169(6):2191-8. doi: 10.1084/jem.169.6.2191.
Polymerase chain reaction has been used to detect increased levels of EBV DNA in salivary gland (SG) biopsies and PBL from patients with Sjogren's syndrome (SS). These results suggest that EBV, which has a normal site of latency in a small number of SG epithelial cells, may be reactivated in SS patients and provide a target for immune attack. The great sensitivity of polymerase chain reaction (PCR) and the ability to analyze very small tissue biopsies (37) make this technique well suited for clinical diagnosis. Specific methods to prevent artefactual contamination of tissue biopsy DNA with viral DNA of other samples (i.e., lyophilization of samples before DNA extraction) and the use of an internal positive control (i.e., inclusion of primers for a single copy human gene) during PCR amplification are presented. Since EBV reactivation occurs with markedly increased frequency in patients with lymphoproliferative and immunodeficiency diseases, as well as transplant recipients receiving cyclosporin A (10), rapid methods of viral detection such as PCR may allow better monitoring of medications and early detection of EBV-related lymphomas that may arise in these patients.
聚合酶链反应已被用于检测干燥综合征(SS)患者唾液腺(SG)活检组织和外周血淋巴细胞(PBL)中EBV DNA水平的升高。这些结果表明,EBV在少数SG上皮细胞中有正常的潜伏位点,在SS患者中可能被重新激活,并成为免疫攻击的靶点。聚合酶链反应(PCR)的高灵敏度以及分析非常小的组织活检样本的能力(37)使该技术非常适合临床诊断。本文介绍了防止组织活检DNA被其他样本的病毒DNA人为污染的具体方法(即DNA提取前对样本进行冻干),以及在PCR扩增过程中使用内部阳性对照(即包含单拷贝人类基因的引物)。由于EBV重新激活在淋巴增殖性疾病、免疫缺陷疾病患者以及接受环孢素A治疗的移植受者中发生的频率显著增加(10),像PCR这样的快速病毒检测方法可能有助于更好地监测药物治疗,并早期发现这些患者可能出现的EBV相关淋巴瘤。