Tsai M M, O'Leary T J
Department of Cellular Pathology, Armed Forces Institute of Pathology, Washington, DC 20306-6000.
Mod Pathol. 1993 Mar;6(2):185-8.
We investigated the use of polymerase chain reaction (PCR) for diagnosis of toxoplasmosis in formalin-fixed, paraffin-embedded tissue, using seven cases in which the diagnosis was certain based on histologic examination and eight cases in which histologic findings only suggested Toxoplasma gondii infection. After amplification the PCR products were subjected to agarose gel electrophoresis followed by Southern blotting with a 32P-labeled oligonucleotide probe. In six of seven proven cases of toxoplasmosis, organisms were detected using DNA extracted from 1/50 to 3/50 of a 6-microns tissue section. In addition, organisms were detected in one of eight cases in which infection was suspected, but not proven by identification of organisms. We conclude on the basis of this study that PCR may be useful for detection of T. gondii in a few cases in which routine histologic examination or immunoperoxidase staining is not definitive. Since PCR has 1000-fold higher sensitivity when applied to fresh, rather than paraffin-embedded, tissues, we expect PCR to be more sensitive in fresh tissue.
我们研究了聚合酶链反应(PCR)在福尔马林固定、石蜡包埋组织中诊断弓形虫病的应用,研究对象包括7例经组织学检查确诊的病例以及8例组织学检查结果仅提示弓形虫感染的病例。PCR产物扩增后进行琼脂糖凝胶电泳,随后用32P标记的寡核苷酸探针进行Southern印迹分析。在7例经证实的弓形虫病病例中,有6例使用从6微米组织切片的1/50至3/50中提取的DNA检测到了病原体。此外,在8例疑似感染但未通过病原体鉴定证实感染的病例中,有1例检测到了病原体。基于这项研究,我们得出结论,在一些常规组织学检查或免疫过氧化物酶染色无法确诊的病例中,PCR可能有助于检测弓形虫。由于PCR应用于新鲜组织而非石蜡包埋组织时灵敏度高1000倍,我们预计PCR在新鲜组织中会更敏感。