A COMPARISON OF TRANSMISSION-BLOCKING ACTIVITY WITH REACTIVITY IN A PLASMODIUM-FALCIPARUM 48/45-KD MOLECULE-SPECIFIC COMPETITION ENZYME-LINKED-IMMUNOSORBENT-ASSAY
A COMPARISON OF TRANSMISSION-BLOCKING ACTIVITY WITH REACTIVITY IN A PLASMODIUM-FALCIPARUM 48/45-KD MOLECULE-SPECIFIC COMPETITION ENZYME-LINKED-IMMUNOSORBENT-ASSAY
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DOI：
10.4269/ajtmh.1995.52.60
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发表时间：
1995-01-01
影响因子：
3.3
通讯作者：
BECKERS, PJA
中科院分区：
文献类型：
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作者：
ROEFFEN, W;LENSEN, T;BECKERS, PJA
单克隆抗体 (MAb) 32F1 和 32F3 与恶性疟原虫 (Pfs48/45) 大配子和受精卵表面表达的分子量分别为 48 和 45 千道尔顿 (kD) 的蛋白质双联体的两个独立表位发生反应；只有 32F3 阻止传输。这些 MAb 用于开发 Pfs48/45 特异性竞争酶联免疫吸附测定 (ELISA)，使用 32F1 捕获抗原并标记 32F3，以定量和分析人血清中抗体对传播阻断活性的贡献。采用比较分析来确定在三组血清样本的生物测定中观察到的竞争 ELISA 滴度和传播阻断活性的一致性：37 份来自曾接触过疟疾的欧洲旅行者，56 份来自配子体携带者，66 份来自喀麦隆疟疾流行地区的学童。配子体携带者和旅行者中 ELISA 和传播阻断测定结果之间的一致性指数被明确定义为中等至中等；对于小学生来说，这种一致性并不显着。所有血清的联合分析显示，竞争性 ELISA 和传播阻断试验的结果之间存在显着且中等至中度的一致性，相对特异性为 94%（传播阻断试验中 105 例阴性，竞争 ELISA 中 99 例也呈阴性），相对敏感性为 44%（传播阻断试验中 54 例阳性，竞争 ELISA 中 24 例也呈阳性）。这项研究表明，C48/45-ELISA 阳性表明蚊子检测中存在传播阻断活性，而阴性结果并不排除传播阻断活性。
Monoclonal antibodies (MAbs) 32F1 and 32F3 react with two independent epitopes of a protein doublet with molecular weights of 48 and 45 kilodaltons (kD) expressed on the surface of Plasmodium falciparum (Pfs48/ 45) macrogametes and zygotes; only 32F3 blocks transmission. These MAbs were used to develop a Pfs48/45-specific competition enzyme-linked immunosorbent assay (ELISA) using 32F1 to capture antigen and labeled 32F3 for quantification and analysis of the contribution of antibodies in human serum to transmission-blocking activity. A comparison analysis was used to determine agreement of competition ELISA titers and transmission-blocking activity as observed in the bioassay in three groups of serum samples: 37 from European travelers with previous exposure to malaria, 56 from gametocyte carriers, and 66 from schoolchildren from a malaria-endemic area in Cameroon. The index of agreement between outcomes of the ELISA and transmission-blocking assay in gametocyte carriers and in travelers was specifically defined as fair-to-moderate; in schoolchildren the agreement was not significant. The combined analysis of all sera showed a significant and fair-to-moderate agreement between the results of the competition ELISA and the transmission-blocking assay, with a relative specificity of 94% (of 105 cases negative in the transmission-blocking assay, 99 were also negative in the competition ELISA) and a relative sensitivity of 44% (of 54 cases positive in the transmission-blocking assay, 24 were also positive in the competition ELISA). This study shows that a positive C48/45-ELISA is indicative for transmission-blocking activity in the mosquito assay, while a negative result does not exclude transmission-blocking activity.