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Identification of parasite DNA in common bile duct stones by PCR and DNA sequencing  

Jang, Ji-Sun (Department of Internal Medicine, Kangdong Sacred Heart Hospital, Hallym University)
Kim, Kyung-Ho (Department of Internal Medicine, Kangdong Sacred Heart Hospital, Hallym University)
Yu, Jae-Ran (Department of Environmental and Tropical Medicine, College of Medicine, Konkuk University)
Lee, Soo-Ung (Department of Environmental and Tropical Medicine, College of Medicine, Konkuk University)
Publication Information
Parasites, Hosts and Diseases / v.45, no.4, 2007 , pp. 301-306 More about this Journal
Abstract
We attempted to identify parasite DNA in the biliary stones of humans via PCR and DNA sequencing. Genomic DNA was isolated from each of 15 common bile duct (CBD) stones and 5 gallbladder (GB) stones. The patients who had the CBD stones suffered from cholangitis, and the patients with GB stones showed acute cholecystitis, respectively. The 28S and 18S rDNA genes were amplified successfully from 3 and/or 1 common bile duct stone samples, and then cloned and sequenced. The 28S and 18S rDNA sequences were highly conserved among isolates. Identity of the obtained 28S D1 rDNA with that of Clonorchis sinensis was higher than 97.6%, and identity of the 18S rDNA with that of other Ascarididae was 97.9%. Almost no intra-specific variations were detected in the 28S and 18S rDNA with the exception of a few nucleotide variations, i.e., substitution and deletion. These findings suggest that C. sinensis and Ascaris lumbricoides may be related with the biliary stoneformation and development.
Keywords
Clonorchis sinensis; Ascaris lumbricoides; ribosomal DNA; biliary stone;
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