Evaluation of loop-mediated isothermal amplification for detection of Toxoplasma gondii in water samples and comparative findings by polymerase chain reaction and immunofluorescence test (IFT)
Evaluation of loop-mediated isothermal amplification for detection of Toxoplasma gondii in water samples and comparative findings by polymerase chain reaction and immunofluorescence test (IFT)
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DOI：
10.1016/j.diagmicrobio.2008.07.009
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发表时间：
2008-12-01
影响因子：
2.9
通讯作者：
Karanis, Panagiotis
中科院分区：
文献类型：
--
作者：
Sotiriadou, Isaia;Karanis, Panagiotis
建立了一种快速检测水样中弓形虫的单管快速环介导等温扩增（LAMP）方法。通过对弓形虫B1和TgOWP基因的扩增，对该方法进行了评价，结果表明，该方法的灵敏度为0.1个速殖子DNA。LAMP检测进行了评估，并与巢式聚合酶链反应（PCR）在26个水样颗粒与已知数量的弓形虫卵囊加标进行了比较。DNA提取后，LAMP和PCR检测的灵敏度分别为100%和53.8%。随后，52个不同来源的天然水样品直接调查3种方法：LAMP，PCR和免疫荧光试验（IFT）。25（48%）的52个已被发现弓形虫DNA阳性的LAMP，而巢式PCR产物中产生的752（13.5%）的水样。所有52份水样经IFT检测均为弓形虫阴性。这些数据清楚地表明，LAMP作为一种快速，特异性和灵敏的工具，用于检测水样中的弓形虫污染。(C)2008年爱思唯尔公司All rights reserved.
The development and evaluation of a l-step single-tube accelerated loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Toxoplasma in water samples is described. The method has been evaluated based on the amplification of B1 and TgOWP Toxoplasma genes, and it demonstrated a sensitivity detection limit of 0.1 tachyzoites' DNA for both genes. LAMP detection was evaluated and compared with nested polymerase chain reaction (PCR) in 26 water sample pellets spiked with known numbers of Toxoplasma oocysts. After DNA extraction, the detection sensitivity in spiked pellets was 100% by LAMP and 53.8% by PCR. Subsequently, 52 natural water samples of different origin were directly investigated by 3 assays: LAMP, PCR, and immunofluorescence test (IFT). Twenty-five (48%) of 52 have been found positive for Toxoplasma DNA by LAMP, whereas nested PCR products were generated in 7 of 52 (13.5%) water samples. All 52 water samples were negative for Toxoplasma by IFT. These data clearly indicate LAMP as a rapid, specific, and sensitive tool for the detection of Toxoplasma contamination in water samples. (C) 2008 Elsevier Inc. All rights reserved.