Direct blood dry LAMP: a rapid, stable, and easy diagnostic tool for Human African Trypanosomiasis.
Direct blood dry LAMP: a rapid, stable, and easy diagnostic tool for Human African Trypanosomiasis.
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DOI：
10.1371/journal.pntd.0003578
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发表时间：
2015-03
影响因子：
3.8
通讯作者：
Sugimoto C
中科院分区：
文献类型：
--
作者：
Hayashida K;Kajino K;Hachaambwa L;Namangala B;Sugimoto C
环介导等温扩增（LAMP）是一种快速、灵敏的工具，可用于多种传染病的诊断。该方法相对于聚合酶链式反应的优点之一是DNA扩增在恒定温度下发生，通常在60-65°C之间;因此，该步骤不需要昂贵的设备。然而，LAMP仍然需要复杂的样品制备步骤和设备齐全的实验室来产生可靠和可重复的结果，这限制了其在大多数发展中国家资源贫乏的实验室中的使用。在这项研究中，我们做了几个实质性的修改技术进行现场诊断的人非洲锥虫病（HAT）在偏远地区使用LAMP。第一个重要的改进是LAMP试剂通过掺入海藻糖作为冷冻保护剂在单管中干燥和稳定，以延长环境温度下的保质期。第二项技术改进是通过简化样品制备步骤实现的，这样可以直接从洗涤剂裂解的血液样品中扩增DNA或RNA。通过这些修改，HAT在流行区的当地诊所或村庄的诊断成为现实，这不仅对HAT而且对其他热带疾病的诊断的应用可能产生重大影响。非洲人锥虫病（HAT）是一种影响许多非洲国家的地方性原生动物病，主要发生在采采蝇病媒存在的农村地区。HAT诊断目前依赖于传统的显微镜技术，其提供相对低的灵敏度，并且通常不能区分两种不同的HAT引起的亚种，T。B. rhodesiense和T. B.冈比亚的。HAT诊断的延迟可能导致疾病从早期（血淋巴）进展到晚期（脑膜脑炎）;因此，快速灵敏的诊断测试对其成功治疗至关重要。在这项研究中，我们开发了一种简单而高度敏感的HAT诊断的分子技术。环介导等温扩增（LAMP）是一种在等温条件下以高灵敏度快速扩增DNA的方法。我们通过稳定试剂和简化样品制备来优化和改进HAT诊断技术。整个程序非常简单，当地工作人员可以在不需要昂贵的设备和专业知识的情况下执行。
Loop-mediated isothermal amplification (LAMP) is a rapid and sensitive tool used for the diagnosis of a variety of infectious diseases. One of the advantages of this method over the polymerase chain reaction is that DNA amplification occurs at a constant temperature, usually between 60–65°C; therefore, expensive devices are unnecessary for this step. However, LAMP still requires complicated sample preparation steps and a well-equipped laboratory to produce reliable and reproducible results, which limits its use in resource-poor laboratories in most developing countries. In this study, we made several substantial modifications to the technique to carry out on-site diagnosis of Human African Trypanosomiasis (HAT) in remote areas using LAMP. The first essential improvement was that LAMP reagents were dried and stabilized in a single tube by incorporating trehalose as a cryoprotectant to prolong shelf life at ambient temperature. The second technical improvement was achieved by simplifying the sample preparation step so that DNA or RNA could be amplified directly from detergent-lysed blood samples. With these modifications, diagnosis of HAT in local clinics or villages in endemic areas becomes a reality, which could greatly impact on the application of diagnosis not only for HAT but also for other tropical diseases. Human African trypanosomiasis (HAT) is an endemic protozoan disease affecting many African countries, predominantly in rural areas where the tsetse fly vector is present. HAT diagnosis currently relies on conventional microscopy techniques, which provide relatively low sensitivity and are often unable to distinguish between the two different HAT-causing subspecies, T. b. rhodesiense and T. b. gambiense. A delay in HAT diagnosis can result in disease progression from an early stage (hemolymphatic) to a late stage (meningoencephalitic); therefore, a rapid and sensitive diagnostic test is crucial to its successful treatment. In this study, we developed a simple and highly sensitive molecular technique for HAT diagnosis. Loop-mediated isothermal amplification (LAMP) is a method that rapidly amplifies DNA with high sensitivity under isothermal conditions. We optimized and modified the technique by stabilizing the reagents and simplifying the sample preparation for the diagnosis of HAT. The whole protocol is so simple that local staff can perform it without the need for costly equipment and expertise.
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影响因子：
3.8
作者：
Mitashi P;Hasker E;Ngoyi DM;Pyana PP;Lejon V;Van der Veken W;Lutumba P;Büscher P;Boelaert M;Deborggraeve S
通讯作者：
Deborggraeve S
DOI：
10.3390/molecules13081773
发表时间：
2008-08-21
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
Teramoto N;Sachinvala ND;Shibata M
通讯作者：
Shibata M
影响因子：
3.2
作者：
Namangala B;Hachaambwa L;Kajino K;Mweene AS;Hayashida K;Simuunza M;Simukoko H;Choongo K;Chansa P;Lakhi S;Moonga L;Chota A;Ndebe J;Nsakashalo-Senkwe M;Chizema E;Kasonka L;Sugimoto C
通讯作者：
Sugimoto C
影响因子：
3.8
作者：
Buescher, Philippe;Ngoyi, Dieudonne Mumba;Bieler, Sylvain
通讯作者：
Bieler, Sylvain
影响因子：
1.5
作者：
BORST, P;VANDERPLOEG, M;JAMES, J
通讯作者：
JAMES, J