Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8842551 | Brazilian Journal of Microbiology | 2018 | 10 Pages |
Abstract
We developed a loop-mediated isothermal amplification (LAMP) assay for the detection of Y. pestis by targeting the 3a sequence on chromosome. All 11 species of the genus Yersinia were used to evaluate the specificity of LAMP and PCR, demonstrating that the primers had a high level of specificity. The sensitivity of LAMP or PCR was 2.3 or 23Â CFU for pure culture, whereas 2.3Â ÃÂ 104 or 2.3Â ÃÂ 106Â CFU for simulated spleen and lung samples. For simulated liver samples, the sensitivity of LAMP was 2.3Â ÃÂ 106Â CFU, but PCR was negative at the level of 2.3Â ÃÂ 107Â CFU. After simulated spleen and lung samples were treated with magnetic beads, the sensitivity of LAMP or PCR was 2.3Â ÃÂ 103 or 2.3Â ÃÂ 106Â CFU, whereas 2.3Â ÃÂ 105 or 2.3Â ÃÂ 107Â CFU for magnetic bead-treated liver samples. These results indicated that some components in the tissues could inhibit LAMP and PCR, and liver tissue samples had a stronger inhibition to LAMP and PCR than spleen and lung tissue samples. LAMP has a higher sensitivity than PCR, and magnetic bead capture of DNAs could remarkably increase the sensitivity of LAMP. LAMP is a simple, rapid and sensitive assay suitable for application in the field or poverty areas.
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Authors
Na Feng, Yazhou Zhou, Yanxiao Fan, Yujing Bi, Ruifu Yang, Yusen Zhou, Xiaoyi Wang,