Polymerase Chain Reaction-Single Strand Confirmation Polymorphism for the Detecti

有许多选项可用于检测和 临床标本中微孢子虫的分型。光学显微镜 允许检测寄生虫，但不允许物种形成。 电子显微镜是物种形成的黄金标准，但 作为一种检测方法不灵敏的。聚合酶链式反应 (聚合酶链式反应)是一种敏感技术，有许多不同的方法 确认阳性结果并确定属和种 导致感染的。物种形成可以通过使用 在聚合酶链式反应分析中或通过使用DNA探针的物种特异性引物 或限制性内切酶扩增后确定物种 进行检测。单链确认多态 (SSCP)与聚合酶链式反应(PCR-SSCP)相结合用于鉴定 细菌、真菌和病毒。我们将使用我们的聚合酶链式反应分析 粪便标本中微孢子虫的检测及SSCP方法的应用 确定寄生虫的具体属和种。生物体 从细胞培养中提取的蛋白质也将用于验证该方法。这 该项目的一部分已接近完成，不久将提交 用于出版。该项目已扩展为将SSCP应用于 流行病学用微孢子虫基因分型的检测 学习。新公布的引物允许物种形成时， 产物用限制性内切酶进行消化。SSCP分析将 使这项任务比限制性内切酶更容易和更敏感 分析。

There are many options for the detection and speciation of microsporidia in clinical specimens. Light microscopy allows for detection of the parasites, but does not allow speciation. Electron microscopy is the gold standard for speciation, but is insensitive as a method of detection. The polymerase chain reaction (PCR) is a sensitive technique with many different methods for confirming a positive result and determining the genus and species causing an infection. Speciation can be achieved by using species-specific primers in the PCR assay, or by using DNA probes or restriction endonucleases to determine the species after the PCR assay is performed.Single strand confirmation polymorphism (SSCP) combined with PCR (PCR-SSCP) has been used to identify bacteria, fungi, and viruses. We will use our PCR assay for the detection of microsporidia in stool specimens and apply SSCP to determine the specific genus and species of the parasite. Organisms from cell culture will also be used to validate the method. This portion of the project is near completion and will soon be submitted for publication.The project has been expanded to apply SSCP to the detection of microsporidial genotypes for use in epidemiological studies. Newly published primers allow speciation when the PCR products are digested with restriction enzymes. SSCP anaylsis will make this task easier and more sensitive than restriction enzyme analysis.