Diagnostic accuracy of loopamp Trypanosoma brucei detection kit for diagnosis of human African trypanosomiasis in clinical samples.
Diagnostic accuracy of loopamp Trypanosoma brucei detection kit for diagnosis of human African trypanosomiasis in clinical samples.
复制标题
DOI：
10.1371/journal.pntd.0002504
复制
发表时间：
2013
影响因子：
3.8
通讯作者：
Deborggraeve S
中科院分区：
文献类型：
--
作者：
Mitashi P;Hasker E;Ngoyi DM;Pyana PP;Lejon V;Van der Veken W;Lutumba P;Büscher P;Boelaert M;Deborggraeve S
分子方法在诊断人类非洲锥虫病（HAT）的敏感寄生虫检测方面具有很大的潜力，但实验室基础设施方面的要求限制了它们在参考中心的使用。最近开发的一种检测方法在等温扩增条件下检测锥虫重复插入移动的元件（RIME）DNA，并已转化为即用型试剂盒形式，即环扩增型布氏锥虫。在本研究中，我们评价了环扩增布氏锥虫检测试剂盒（以下称为LAMP）在确诊结核病中的诊断性能。来自刚果民主共和国（DRC）的冈比亚HAT患者、HAT疑似者和健康地方病控制者。142 T. b.冈比亚HAT患者，111名健康的地方病对照和97名未经证实的HAT可疑者被纳入这项回顾性评价。参考标准检测是血液、淋巴液或脑脊液中的寄生虫检测。用LAMP一式两份地分析来自所有研究参与者的血液的存档DNA。LAMP在寄生虫学确诊病例中的灵敏度在第一轮为87.3%（95%CI 80.9-91.8%），在第二轮为93.0%（95%CI 87.5-96.1%）。健康对照的特异性在第一次运行中为92.8%（95% CI 86.4-96.3%），在第二次运行中为96.4%（95% CI 91.1-98.6%）。再现性极好，Kappa值为0.81。在这项基于实验室的研究中，Loopamp布氏锥虫检测试剂盒显示出良好的诊断准确性和良好的重现性。需要进一步的研究，以评估其常规使用的可行性，在现场条件下诊断HAT。诊断和有效治疗是控制人类非洲锥虫病（HAT）的基石。聚合酶链反应（PCR）等分子工具检测寄生虫的DNA，通常非常敏感和特异。然而，PCR不适用于现场环境，因为它需要实验室基础设施和复杂的设备。最近开发的环介导等温扩增（LAMP）已成为一个更简单的替代传统的分子方法诊断HAT。该检测试剂盒已被转化为用于定性检测临床标本中寄生虫DNA的诊断试剂盒，即Loopamp Trypanosoma brucei Detection Kit。在这项研究中，我们在实验室条件下对从刚果民主共和国的142名患者、97名嫌疑人和111名健康地方病控制者的血液样本中提取的DNA进行了评估。该试验具有良好的诊断准确性和重现性。鉴于LAMP相对于常规核酸方法的实际优势，这些是有希望的结果。需要进一步的研究来评估测试的准确性和在现场条件下的可行性。
Molecular methods have great potential for sensitive parasite detection in the diagnosis of human African trypanosomiasis (HAT), but the requirements in terms of laboratory infrastructure limit their use to reference centres. A recently developed assay detects the Trypanozoon repetitive insertion mobile element (RIME) DNA under isothermal amplification conditions and has been transformed into a ready-to-use kit format, the Loopamp Trypanosoma brucei. In this study, we have evaluated the diagnostic performance of the Loopamp Trypanosoma brucei assay (hereafter called LAMP) in confirmed T.b. gambiense HAT patients, HAT suspects and healthy endemic controls from the Democratic Republic of the Congo (DRC). 142 T.b. gambiense HAT patients, 111 healthy endemic controls and 97 HAT suspects with unconfirmed status were included in this retrospective evaluation. Reference standard tests were parasite detection in blood, lymph or cerebrospinal fluid. Archived DNA from blood of all study participants was analysed in duplicate with LAMP. Sensitivity of LAMP in parasitologically confirmed cases was 87.3% (95% CI 80.9–91.8%) in the first run and 93.0% (95% CI 87.5–96.1%) in the second run. Specificity in healthy controls was 92.8% (95% CI 86.4–96.3%) in the first run and 96.4% (95% CI 91.1–98.6%) in the second run. Reproducibility was excellent with a kappa value of 0.81. In this laboratory-based study, the Loopamp Trypanosoma brucei Detection Kit showed good diagnostic accuracy and excellent reproducibility. Further studies are needed to assess the feasibility of its routine use for diagnosis of HAT under field conditions. Diagnosis and effective treatment are cornerstones in the control of human African trypanosomiasis (HAT). Molecular tools such as the polymerase chain reaction (PCR) detect the parasite's DNA and are generally very sensitive and specific. However, PCR is not applicable in field settings because it requires a laboratory infrastructure and sophisticated equipment. A recently developed loop-mediated isothermal amplification (LAMP) has emerged as a simpler alternative to conventional molecular methods for the diagnosis of HAT. The test has been transformed into a diagnostic kit for qualitative detection of the parasite's DNA in clinical specimens, the Loopamp Trypanosoma brucei Detection Kit. In this study, we evaluated this kit in laboratory conditions on DNA extracted from blood samples of 142 patients, 97 suspects and 111 healthy endemic controls in the Democratic Republic of the Congo. The test showed good diagnostic accuracy and excellent reproducibility. Given the practical advantages of LAMP over conventional nucleic acid methods these are promising results. Further studies are needed to assess the test's accuracy and feasibility in field conditions.
登录
查看更多内容
影响因子：
3.8
作者：
Hasker E;Lutumba P;Chappuis F;Kande V;Potet J;De Weggheleire A;Kambo C;Depoortere E;Pécoul B;Boelaert M
通讯作者：
Boelaert M
影响因子：
3.8
作者：
Deborggraeve, Stijn;Lejon, Veerle;Buscher, Philippe
通讯作者：
Buscher, Philippe
影响因子：
2.7
作者：
Solano, P;Jamonneau, V;Garcia, A
通讯作者：
Garcia, A
影响因子：
15.8
作者：
Simarro, Pere P.;Jannin, Jean;Cattand, Pierre
通讯作者：
Cattand, Pierre
影响因子：
3.8
作者：
Buescher, Philippe;Ngoyi, Dieudonne Mumba;Bieler, Sylvain
通讯作者：
Bieler, Sylvain