Immunocapture diagnostic assays for malaria using Plasmodium lactate dehydrogenase (pLDH)
Immunocapture diagnostic assays for malaria using Plasmodium lactate dehydrogenase (pLDH)
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DOI：
10.4269/ajtmh.1999.60.109
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发表时间：
1999-01-01
影响因子：
3.3
通讯作者：
Makler, M
中科院分区：
文献类型：
--
作者：
Piper, R;Lebras, J;Makler, M
我们已经开发了两种诊断方法的基础上特异性检测疟原虫乳酸脱氢酶（pLDH）活性。这些检测利用一组单克隆抗体，捕获寄生虫酶，并允许对人类疟疾感染进行定量和物种形成。免疫捕获pLDH活性测定（ICPLDH）允许使用NAD类似物APAD从感染的血液中快速纯化和测量pLDH，所述NAD类似物APAD与疟原虫LDH同种型特异性反应。还设计了免疫层析试验（OptiMAL（R）测定），并允许检测类似于200个寄生虫/μ l血液的寄生虫感染。通过使用抗体的组合，这两种测试不仅可以检测，而且可以区分恶性疟原虫和非恶性疟原虫疟疾。这两种检测方法的灵敏度与其他商业非显微镜检测方法相当;重要的是，我们发现很少有假阳性样本，特别是最近清除疟疾感染的患者样本。此外，我们发现，当使用定量ICPpLDH测定时，pLDH活性水平密切反映了初始诊断和患者治疗后的寄生虫血症水平。我们的结论是，诊断试验的基础上检测pLDH的检测是敏感和实用的检测，形态，和定量的所有人类疟原虫感染，也可以用来指示耐药感染。
We have developed two diagnostic assays based on the specific detection of Plasmodium lactate dehydrogenase (pLDH) activity. These assays exploit a panel of monoclonal antibodies that capture the parasite enzyme and allow for the quantitation and speciation of human malaria infections. An immunocapture pLDH activity assay (ICpLDH) allows for the rapid purification and measurement of pLDH from infected blood using the NAD analog APAD, which reacts specifically with Plasmodium LDH isoforms. An immunochromatographic test (the OptiMAL(R) assay) was also formatted and allowed the detection of parasite infections of similar to 200 parasites/yl of blood. By using a combination of antibodies, both tests can not only detect but differentiate between P. falciparum and non-P. falciparum malaria. Both assays show a sensitivity comparable with other commercial nonmicroscopic tests; importantly, we found very few instances of false-positive samples, especially with samples from patients recently cleared of malaria infection. Furthermore, we find that when one uses the quantitative ICpLDH assay, the levels of pLDH activity closely mirror the levels of parasitemia in both initial diagnosis and while following patient therapy. We conclude that diagnostic tests based on the detection of pLDH are both sensitive and practical for the detection, speciation, and quantitation of all human Plasmodium infections and can also be used to indicate drug-resistant infections.