80070096

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コウソ ヒョウシキ ゴウセイ オリゴヌクレオチド プローブ ニヨル CAMPYLOBACTER JEJUNI/COLI ノ ジンソク ケンシュツ

The detection of Campylobacter jejuni/coli with the enrichment culture by the enzyme labeled synthetic oligonucleotide probe

Japanese (日本語)

神戸大学医療技術短期大学部紀要

5

37-41

神戸大学医療技術短期大学部

1989

2007-11-21

The synthetic oligonucleotide probe labeled by alkalinephosphatase is useful to detect the pure cultured bacteria. In this paper, the probe was used to detect C. jejuni /coli in feces for rapid diagnosis. Direct detection from feces was difficult. Therefore, feces were cultured in the enrichment medium (Hanninen medium) for 6, 20 and 44 hours. Then, the cultured medium was treated with the following methods. 1) Direct method : the cultured medium was directly blotted onto nylon membrane. 2) Centrifugation method : bacteria in the cultured medium was collected by the centrifugation and blotted onto nylon membrane. 3) Phenol extract method : DNA was extracted from the cultured medium by the conventional phenol extract method and blotted onto nylon membrane. And all of these blotted nylon membrane were used for hybridization. This probe was applied to detect to C. jejuni/coli in 76 clinical specimens (feces in Cary-Blair medium). Among them, 6 specimens were proved positive in 24 hours, though the conventional system took 2-3 days to prove it. The phenol extract method was more sensitivity than other two methods, but some data were false positive. Concerning clinical specimens, the centrifugation method was more sensitive than the direct method and more specific than the phenol extract method. Consequently, the centrifugation method was simple in process and satisfactory to detect C. jejuni/coli from the cultured medium.

紀要論文