Development of loop-mediated isothermal amplification (LAMP) method for diagnosis of equine piroplasmosis
Development of loop-mediated isothermal amplification (LAMP) method for diagnosis of equine piroplasmosis
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DOI：
10.1016/j.vetpar.2006.08.014
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发表时间：
2007-01-31
影响因子：
2.6
通讯作者：
Igarashi, Ikuo
中科院分区：
文献类型：
--
作者：
Alhassan, Andy;Thekisoe, Oriel M. M.;Igarashi, Ikuo
环介导等温扩增 (LAMP) 是一种新型核酸方法，使用一组四个专门设计的引物和具有链置换活性的 DNA 聚合酶，在等温条件下以高特异性、高效和快速的方式扩增 DNA。在本研究中，我们使用根据 EMA-1 和 Bc 48 基因设计的 LAMP 引物组分别检测马泰勒虫和卡巴贝虫感染。这些引物组特异性扩增了相应寄生虫的 DNA。两个引物组均将 T equal 和 B. caballi 扩增至 10 倍连续稀释样品的 10(-6) 稀释度。此外，在感染后 2 至 35 天，从实验性感染 T equal 的马血液中提取的 DNA 通过 T equal LAMP 引物组进行扩增，证明了这些引物的高灵敏度。在从中国收集的 55 个样本中，LAMP 检测结果分别为马 T 马和马巴利布鲁氏菌感染的阳性率为 81.8% 和 56.3%。相比之下，从南非采集的 37 个样本中，马齿苋和 B. caballi 的 LAMP 阳性率分别为 91.8% 和 45.9%。这些结果表明，LAMP 可能成为马梨形虫病流行病学研究的潜在诊断工具。 (c) 2006 Elsevier B.V. 保留所有权利。
Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid method whereby DNA is amplified with high specificity, efficiency, and rapidity under isothermal conditions using a set of four specifically designed primers and a DNA polymerase with strand displacement activity. In this study, we used LAMP primer sets designed from EMA-1 and Bc 48 genes for detection of Theileria equi and Babesia caballi infections, respectively. These primer sets specifically amplified DNA of the respective parasites. Both primer sets amplified T equi and B. caballi up to 10(-6) dilution of 10-fold serially diluted samples. Furthermore, DNA extracted from blood collected from a horse experimentally infected with T equi was amplified by a T equi LAMP primer set from days 2 to 35 post-infection, demonstrating the high sensitivity of these primers. Of 55 samples collected from China, 81.8% and 56.3% were positively detected by LAMP for T equi and B. caballi infections, respectively. In contrast, 91.8% and 45.9% of the 37 samples collected from South Africa were LAMP positive for T equi and B. caballi, respectively. These results suggest that LAMP could be a potential diagnostic tool for epidemiological studies of equine piroplasmosis. (c) 2006 Elsevier B.V. All rights reserved.