著者

CAO Lili CHENG Ronghua YAO Lin YUAN Shuxian YAO Xinhua

出版者

公益社団法人 日本獣医学会

雑誌

The Journal of Veterinary Medical Science (ISSN:09167250)

巻号頁・発行日

vol.76, no.1, pp.9-14, 2014

被引用文献数

10

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The Loop-mediated isothermal amplification (LAMP) method amplifies DNA with high simply, specificity, sensitivity and rapidity. In this study, A LAMP assay with 6 primers targeting a highly conserved region of the GRA1 gene was developed to diagnose <i>Toxoplasma gondii</i>. The reaction time of the LAMP assay was shortened to 30 min after optimizing the reaction system. The LAMP assay was found to be highly specific and stable. The detection limit of the LAMP assay was 10 copies, the same as that of the conventional PCR. We used the LAMP assay to develop a real-time fluorogenic protocol to quantitate <i>T. gondii</i> DNA and generated a log-linear regression plot by plotting the time-to-threshold values against genomic equivalent copies. Furthermore, the LAMP assay was applied to detect <i>T. gondii</i> DNA in 423 blood samples and 380 lymph node samples from 10 pig farms, and positive results were obtained for 7.8% and 8.2% of samples, respectively. The results showed that the LAMP method is slightly more sensitive than conventional PCR (6.1% and 7.6%). Positive samples obtained from 6 pig farms. The LAMP assay established in this study resulted in simple, specific, sensitive and rapid detection of <i>T. gondii</i> DNA and is expected to play an important role in clinical detection of <i>T. gondii</i>.