Querol J M, Farga A, Alonso C, Granda D, Alcaraz M J, García de Lomas J
Servicio de Medicina Interna, Hospital Lluìs Alcanyis, Xativa.
An Med Interna. 1996 May;13(5):235-8.
The utility of polymerase chain reaction (PCR) is described for the diagnosis in three patients suffering from central nervous system infections, tuberculous meningitis, herpetic encephalitis and cerebral toxoplasmosis. PCR was performed in the cerebrospinal fluid after processing the specimen by two methods, proteinase K digestion and phenol extraction of DNA. Amplification was realized using primers previously described that amplify specific DNA fragments of each microorganisms (insertion sequence IS6110 of Mycobacterium tuberculosis, B1 gene of Toxoplasma gondii, and DNA polymerase gene of Herpes simplex virus). In all three cases, PCR was positive after amplification of the specimen extracted with proteinase K, as well as when a complete DNA extraction with phenol was realized. In all cases a band of amplified products was observed in agarose gels. In conclusion, in all three patients described, PCR would had allowed the diagnosis in seven hours, and PCR should be consider a rapid sensitive and relatively simple method.
本文描述了聚合酶链反应(PCR)在诊断3例中枢神经系统感染患者中的应用,这些患者分别患有结核性脑膜炎、疱疹性脑炎和脑弓形虫病。通过蛋白酶K消化和DNA苯酚抽提这两种方法处理脑脊液标本后进行PCR检测。使用先前描述的引物实现扩增,这些引物可扩增每种微生物的特定DNA片段(结核分枝杆菌的插入序列IS6110、弓形虫的B1基因和单纯疱疹病毒的DNA聚合酶基因)。在所有3例病例中,用蛋白酶K提取的标本扩增后以及用苯酚进行完全DNA提取时,PCR均呈阳性。在所有病例中,在琼脂糖凝胶中均观察到扩增产物条带。总之,在所描述的所有3例患者中,PCR可在7小时内做出诊断,PCR应被视为一种快速、灵敏且相对简单的方法。