Diagnosis of Sudden death by in-situ-PCR using tissues from Autopsy

本研究是通过检测一种已使用的细菌引物来修复原位PCR的实验条件的。作为一个结果，最好的实验条件可以是固定的，并应用于实际工作。在目前的研究中，我试着检测肺结核样本中细菌感染的存在，并使用DIG寡核苷酸3'-末端标签工具包（Boehringer Mannheim）。我可以确信一个阶段的肺炎在时间上会有所不同，这些方法是通过这些程序提供的，但在那之后，它确定了检测病毒、特别是疱疹组的实验条件，也是如此。此外，它还通过原位RT· PCR对hypoxia条件进行了清单。我检测到了一些细胞因子，例如，红细胞生成素、白细胞介素-1a，并添加到血管内皮素细胞生长因子。However，这些说明在诊断的自动样本中并不太强，并在实际使用中找到了困难。具体地说，从抑郁中突然死亡，对这些标记者来说是一个非常小的描述。因此，要做实际应用，我扩展了一个搜索范围，以获得离细胞反应更近的距离，并找到了协同分析下一步的必要性。

This research did the, detection which used peculiar primer of the bacteria to fix the experimental conditions of in-situ-PCR. As a result, the best experimental conditions could be fixed, and applied for practical work. In the present study, I tried to detect the presence of bacterial infections in lung tissue samples, using a DIG Oligonucleotide 3'-End Labeling Kit (Boehringer Mannheim). I was able to confirm a stage of pneumonia to differ in time course, and high sensitivity of these methods was proved by these procedure.After that, it fixed experimental conditions detectable for virus, especially, the herpes group, too. Also, it detected the RNA which is manifested in the hypoxia condition by the in-situ-RT ・ PCR. I detected some kind of cytokines, for example, erythropoietin, interleukin-1a, in addition to the vascular endothelium cell growth factor.However, these manifestations were not too strong in an autopsy sample to diagnosis, and found the difficulty for practical use. Particularly, sudden death from suffocation, there is a extremely little manifestation of these markers.Therefore, to do practicing application, I expanded a search range for mitochondria DNA having relevance to be close to cellular respiration, and found necessity to analyze synthetically for the next step.