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Reference genes for quantitative analysis on Clonorchis sinensis gene expression by real-time PCR

Cited 58 time in Web of Science Cited 62 time in Scopus
Authors

Yoo, Won Gi; Kim, Tae Im; Li, Shunyu; Kwon, Oh Sil; Cho, Pyo Yun; Kim, Tong-Soo; Kim, Kijeong; Hong, Sung-Jong

Issue Date
2009-01
Publisher
Springer Verlag
Citation
Parasitology Research, Vol.104 No.2, pp.321-328
Abstract
The accuracies of relative gene expressions as determined by quantitative real-time polymerase chain reaction are largely dependent on the variabilities of the reference genes used. Validation of the stabilities of reference genes under experimental conditions is an essential initial step for comparative studies on the expression levels of target genes in experimental groups. Using three total RNA samples extracted independently from Clonorchis sinensis metacercariae and adults, we determined the gene expression stabilities of eight reference gene candidates and the relative transcript levels of three target genes using the geNorm program. The reference genes found to be stably expressed in metacercariae and adults were phosphoglycerate kinase, beta-actin, and calcyphosine; reference genes found to be stably expressed under gamma-irradiated and non-irradiated conditions were succinate dehydrogenase, small nuclear ribonucleoprotein, and beta-actin; and those stably expressed regardless of bile treatment were small nuclear ribonucleoprotein, phosphoglycerate kinase, and succinate dehydrogenase. According to our data, the expression levels of target genes are dependent on normalization factors, such as the C (T) values of single reference genes and the geometric mean of the C (T) values of three reference genes. When comparing C. sinensis gene expressions, we propose to employ the geometric mean of the C (T) values of more than three reference genes validated in the same experimental setting.
ISSN
0932-0113
URI
https://hdl.handle.net/10371/200705
DOI
https://doi.org/10.1007/s00436-008-1195-x
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  • College of Veterinary Medicine
  • Department of Veterinary Medicine
Research Area Microbiology, Parasitology, Tropical Medicine

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