Loop-mediated isothermal amplification (LAMP): early detection of Toxoplasma gondii infection in mice.
Loop-mediated isothermal amplification (LAMP): early detection of Toxoplasma gondii infection in mice.
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DOI：
10.1186/1756-3305-5-2
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发表时间：
2012-01-03
影响因子：
3.2
通讯作者：
Zhou XN
中科院分区：
文献类型：
--
作者：
Kong QM;Lu SH;Tong QB;Lou D;Chen R;Zheng B;Kumagai T;Wen LY;Ohta N;Zhou XN
弓形虫病是一种广泛存在于动物和人类中的人畜共患寄生虫病。传统的分子检测方法往往难以进行，特别是对于弓形虫感染的早期诊断。我们建立了一种新的以529 bp重复序列为靶点的环介导等温扩增技术（529 bp-LAMP）。RH株速殖子感染实验小鼠血样中弓形虫DNA的含量。该测定用Bst DNA聚合酶在65°C下进行1小时。529 bp-LAMP方法的检测限低至0.6fg。弓形虫DNA该方法的灵敏度分别比靶向B1基因的LAMP（B1-LAMP）和靶向529 bp重复序列的巢式PCR（529 bp-巢式PCR）高100倍和1000倍。用伊氏锥虫、恶性疟原虫、卫氏并殖吸虫、日本血吸虫、肝片吸虫和广州管圆线虫的DNA样品测定529 bp-LAMP法的特异性。该试剂盒可检测T。在感染后一天（dpi）所有小鼠血液样品中的弓形虫DNA。结果表明：（1）529 bp-LAMP法对T的检出限为0.6fg;弓形虫DNA;（ii）该检测不涉及与其他寄生虫的DNA的任何交叉反应性;（iii）这是关于应用LAMP检测对来自实验感染小鼠的血液样品中的弓形虫病进行早期诊断的第一份报告。由于其简单，敏感性和成本效益的共同使用，我们建议，该检测应作为一个早期诊断工具，弓形虫病的健康控制。
Toxoplasmosis is a widespread zoonotic parasitic disease that occurs in both animals and humans. Traditional molecular assays are often difficult to perform, especially for the early diagnosis of Toxoplasma gondii infections. Here, we established a novel loop-mediated isothermal amplification targeting the 529 bp repeat element (529 bp-LAMP) to detect T. gondii DNA in blood samples of experimental mice infected with tachyzoites of the RH strain. The assay was performed with Bst DNA polymerase at 65°C for 1 h. The detection limit of the 529 bp-LAMP assay was as low as 0.6 fg of T. gondii DNA. The sensitivity of this assay was 100 and 1000 fold higher than that of the LAMP targeting B1 gene (B1-LAMP) and nested PCR targeting 529 bp repeat element (529 bp-nested PCR), respectively. The specificity of the 529 bp-LAMP assay was determined using the DNA samples of Trypanosoma evansi, Plasmodium falciparum, Paragonimus westermani, Schistosoma japonicum, Fasciola hepatica and Angiostrongylus cantonensis. No cross-reactivity with the DNA of any parasites was found. The assay was able to detect T. gondii DNA in all mouse blood samples at one day post infection (dpi). We report the following findings: (i) The detection limit of the 529 bp-LAMP assay is 0.6 fg of T. gondii DNA; (ii) The assay does not involve any cross-reactivity with the DNA of other parasites; (iii) This is the first report on the application of the LAMP assay for early diagnosis of toxoplasmosis in blood samples from experimentally infected mice. Due to its simplicity, sensitivity and cost-effectiveness for common use, we suggest that this assay should be used as an early diagnostic tool for health control of toxoplasmosis.
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影响因子：
3.2
作者：
Abu-Madi, Marawan A.;Al-Molawi, Naema;Behnke, Jerzy M.
通讯作者：
Behnke, Jerzy M.
影响因子：
2.4
作者：
Montoya, Ana;Miro, Guadalupe;Fuentes, Isabel
通讯作者：
Fuentes, Isabel
影响因子：
3.2
作者：
Aonuma H;Yoshimura A;Perera N;Shinzawa N;Bando H;Oshiro S;Nelson B;Fukumoto S;Kanuka H
通讯作者：
Kanuka H
影响因子：
3.3
作者：
Selvey, S;Thompson, EW;Griffiths, LR
通讯作者：
Griffiths, LR
DOI：
10.1016/j.diagmicrobio.2008.07.009
发表时间：
2008-12-01
影响因子：
2.9
作者：
Sotiriadou, Isaia;Karanis, Panagiotis
通讯作者：
Karanis, Panagiotis