Development and comparative evaluation of two antigen detection tests for Visceral Leishmaniasis.
Development and comparative evaluation of two antigen detection tests for Visceral Leishmaniasis.
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DOI：
10.1186/s12879-015-1125-3
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发表时间：
2015-09-22
影响因子：
3.7
通讯作者：
Reed SG
中科院分区：
文献类型：
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作者：
Vallur AC;Tutterrow YL;Mohamath R;Pattabhi S;Hailu A;Abdoun AO;Ahmed AE;Mukhtar M;Salam MA;Almeida ML;Almeida RP;Mondal D;Albertini A;Ghalib H;Duthie MS;Reed SG
如果不及时诊断和治疗，内脏利什曼病（VL）可能是致命的。治疗效果因耐药、药物毒性和合并症而异。重要的是监测治疗反应，以确认治愈和减少复发。目前，脾脏、骨髓或淋巴结活检的显微镜检查是评估治愈的唯一确定方法。对于VL的治疗来说，微创检查是治疗成功的重中之重。在这项研究中，我们描述了基于检测尿液样本中多诺瓦利什曼原虫抗原的捕获ELISA的发展，并与利什曼原虫抗原ELISA的比较，也是为了同样的目的而开发的。这两种试剂盒都是作为原型开发的，并对来自苏丹、埃塞俄比亚、孟加拉国和巴西的患者尿液样本以及来自流行和非流行地区的适当对照样本进行了测试。与对照样本相比，基于准确检测患者的敏感性和特异性进行评估。采用单因素方差分析（One- Way ANOVA）评估测试的辨别能力，并采用Cohen’s kappa评估其相关性。利什曼原虫抗原检测™ELISA对来自苏丹、孟加拉国和埃塞俄比亚的VL患者样本的敏感性为90%，对巴西样本的敏感性为88%。利什曼原虫抗原ELISA除对苏丹样品敏感性较低外，在性能上具有可比性。两者都非常具体。为了确认在监测治疗中的效用，在治疗后0,30和180天收集了VL患者的尿液样本。对于利什曼原虫抗原检测™ELISA，第0天的阳性率高达95%，第30天降至21%。第180天，所有样品均为阴性，符合临床治愈。类似的趋势也见于利什曼原虫抗原ELISA；第0天的阳性率较低，为91%，更多的患者在第30天和180天仍呈阳性。利什曼原虫抗原检测™和利什曼原虫抗原elisa是标准化的、用户友好的、定量的和直接的测试，用于检测急性VL期间的利什曼原虫，以及监测治疗期间的寄生虫清除。它们是对现有选项的明显改进。elisa提供了一种在急性感染期间检测寄生虫抗原并在治愈后监测其清除的非侵入性方法，填补了VL管理中未满足的需求。采用更多来自流行地区的样本进一步改进检测方法，将确定其在监测治疗方面的效用。
Visceral leishmaniasis (VL) can be fatal without timely diagnosis and treatment. Treatment efficacies vary due to drug resistance, drug toxicity and co-morbidities. It is important to monitor treatment responsiveness to confirm cure and curtail relapse. Currently, microscopy of spleen, bone marrow or lymph node biopsies is the only definitive method to evaluate cure. A less invasive test for treatment success is a high priority for VL management. In this study, we describe the development of a capture ELISA based on detecting Leishmania donovani antigens in urine samples and comparison with the Leishmania Antigen ELISA, also developed for the same purpose. Both were developed as prototype kits and tested on patient urine samples from Sudan, Ethiopia, Bangladesh and Brazil, along with appropriate control samples from endemic and non-endemic regions. Sensitivity and specificity were assessed based on accurate detection of patients compared to control samples. One- Way ANOVA was used to assess the discrimination capacity of the tests and Cohen’s kappa was used to assess their correlation. The Leishmania Antigen Detect™ ELISA demonstrated >90 % sensitivity on VL patient samples from Sudan, Bangladesh and Ethiopia and 88 % on samples from Brazil. The Leishmania Antigen ELISA was comparable in performance except for lower sensitivity on Sudanese samples. Both were highly specific. To confirm utility in monitoring treatment, urine samples were collected from VL patients at days 0, 30 and 180 post- treatment. For the Leishmania Antigen Detect™ ELISA, positivity was high at day 0 at 95 %, falling to 21 % at day 30. At day 180, all samples were negative, corresponding well with clinical cure. A similar trend was also seen for the Leishmania Antigen ELISA albeit; with lower positivity of 91 % at Day 0 and more patients, remaining positive at Days 30 and 180. The Leishmania Antigen Detect™ and the Leishmania Antigen ELISAs are standardized, user- friendly, quantitative and direct tests to detect Leishmania during acute VL as well as to monitor parasite clearance during treatment. They are a clear improvement over existing options. The ELISAs provide a non-invasive method to detect parasite antigens during acute infection and monitor its clearance upon cure, filling an unmet need in VL management. Further refinement of the tests with more samples from endemic regions will define their utility in monitoring treatment.