To develop a technique for detecting the fbsB gene of Streptococcus agalactiae using a novel DNA amplification method designated Loop-Mediated Isothermal Amplification (LAMP).The detection assay is based on the loop-mediated isothermal amplification (LAMP) reaction.The fbsB gene was amplified by a set of four specially primers that recognize six distinct sequences of the target.The amplification can be obtained in 1h by incubating all of the reagents in a single tube with Bst DNA polymerase at 63℃.Results showed that the developed LAMP assay demonstrated an exceptionally higher than the conventional PCR.This assay results was found to be 100 times more sensitive than the PCR assay,and consistent with the results of real-time PCR.Our results clearly demonstrate that the LAMP-based assay is a sensitive and reliable means for the detection of Streptococcus agalactiae.
WANG Yong
,
ZHAO Xin
,
JING Hai-chun
,
LAN Qing-kuo
,
ZHU Zhu
,
CHENG Yi
. Development and Application of Loop-Mediated Isothermal Amplification for Detection of Streptococcus agalactiae[J]. Acta Agriculturae Boreali-Sinica, 2009
, 24(5)
: 234
-238
.
DOI: 10.7668/hbnxb.2009.05.050
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