2001 Fiscal Year Final Research Report Summary
Diagnosis of Sudden death by in-situ-PCR using tissues from Autopsy
| Project/Area Number |
11670415
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Legal medicine
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| Research Institution | University of Tsukuba (2001)
Osaka University (1999-2000) |
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Principal Investigator |
HONDA Katsuya University of Tsukuba, Institute of Community of Medicine, Professor, 社会医学系, 教授 (00240789)
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| Project Period (FY) |
1999 – 2001
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| Keywords | in-situ-PCR / Direct method / hypoxia / Pneumonia / mitochondorial DNA |
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| Research Abstract |
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.
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