Preliminary study on detection of Echinococcus granulosus DNA by loop-mediated isothermal amplification

To evaluate the sensitivity of loop-mediated isothermal amplification (LAMP) on the detection of Echinococcus granulosus. The DNAs were extracted from Echinococcus granulosus eggs and adults. According to Echinococcus mitochondrial 12S rRNA sequences and the mechanism of LAMP, 4 Echinococcus specifi...

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Published inZhongguo xue xi chong bing fang zhi za zhi Vol. 23; no. 5; p. 558
Main Authors Xiang-Zhen, Xu, Xiao-Lin, Jin, Jian, Li, Wen-Cai, Jiang, Gang, Jiang
Format Journal Article
LanguageChinese
Published China 01.10.2011
Subjects
Animals
DNA, Helminth - analysis
Echinococcus granulosus - genetics
Nucleic Acid Amplification Techniques - methods
Sensitivity and Specificity
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Summary:To evaluate the sensitivity of loop-mediated isothermal amplification (LAMP) on the detection of Echinococcus granulosus. The DNAs were extracted from Echinococcus granulosus eggs and adults. According to Echinococcus mitochondrial 12S rRNA sequences and the mechanism of LAMP, 4 Echinococcus specific primers were designed and used for LAMP assay, and Bubble taenia and the blank were used as the negative control for evaluation of the specificity. The LAMP products were stained by SYBR Green I and analyzed by electrophoresis, and 1000, 100, 10, 1 eggs of Echinococcus granulosus per 200 microl were amplified by LAMP for evaluating the sensitivity. The LAMP products of Echinococcus granulosus adult DNA became turbid and green after staining while the products of control DNA kept clarify and brown after staining. Electrophoresis analysis showed that the LAMP products of Echinococcus granulosus eggs presented characteristic ladders, but the products of control did not. The detection limit of LAMP assay was 1 egg of
ISSN:1005-6661