Screening for Pfhrp2/3-Deleted Plasmodium falciparum, Non-falciparum, and Low-Density Malaria Infections by a Multiplex Antigen Assay
Screening for Pfhrp2/3-Deleted Plasmodium falciparum, Non-falciparum, and Low-Density Malaria Infections by a Multiplex Antigen Assay
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DOI：
10.1093/infdis/jiy525
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发表时间：
2019-02-01
影响因子：
6.4
通讯作者：
Rogier, Eric
中科院分区：
文献类型：
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作者：
Plucinski, Mateusz M.;Herman, Camelia;Rogier, Eric
背景。疟原虫抗原的检测提供了疟疾感染状况的证据，是大多数疟疾诊断的基础。我们开发了一种灵敏的基于球头的多重检测方法，用于实验室使用，可同时检测泛疟原虫醛缩酶（pAldo）、泛疟原虫乳酸脱氢酶（pLDH）和恶性疟原虫组氨酸富蛋白2 （PfHRP2）抗原。该检测方法在纯化重组抗原、单种疟疾感染和非感染血液样本中得到验证。为了对照在流行环境中收集的样本进行测试，对安哥拉门诊样本（n = 1267）进行了分析。在安哥拉466份至少1种抗原阳性的样本中，最常见的抗原谱为PfHRP2+/pAldo+/pLDH+(167, 36%)、PfHRP2+/pAldo/pLDH（163, 35%）和PfHRP2+/pAldo+/pLDH（129, 28%）。抗原谱预测聚合酶链反应（PCR）阳性和寄生虫密度。8份安哥拉样本（1.7%）没有或极低的PfHRP2，但其他一种或两种抗原呈阳性。PCR分析证实3例（0.6%）为卵形疟原虫感染，2例（0.4%）为缺乏Pfhrp2和/或Pfhrp3的恶性疟原虫。这是安哥拉首次报道Pfhrp2/3缺失突变体。高通量多重抗原检测可以廉价地筛选低密度恶性疟原虫、非恶性疟原虫和pfhrp2 /3缺失寄生虫，以提供种群水平的抗原估计，并确定需要进一步分子表征的标本。建立了一种多重疟疾抗原检测方法，用于检测泛疟原虫抗原醛缩酶、LDH和恶性疟原虫抗原HRP2。多重抗原检测可用于预测疟疾感染状况、感染种类和pfhrp2 /3缺失寄生虫的检测。
Background. Detection of Plasmodium antigens provides evidence of malaria infection status and is the basis for most malaria diagnosis.We developed a sensitive bead-based multiplex assay for laboratory use, which simultaneously detects pan-Plasmodium aldolase (pAldo), pan-Plasmodium lactate dehydrogenase (pLDH), and P. falciparum histidine-rich protein 2 (PfHRP2) antigens. The assay was validated against purified recombinant antigens, monospecies malaria infections, and noninfected blood samples. To test against samples collected in an endemic setting, Angolan outpatient samples (n = 1267) were assayed.Of 466 Angolan samples positive for at least 1 antigen, the most common antigen profiles were PfHRP2+/pAldo+/pLDH+ (167, 36%), PfHRP2+/pAldo/pLDH (163, 35%), and PfHRP2+/pAldo+/pLDH (129, 28%). Antigen profile was predictive of polymerase chain reaction (PCR) positivity and parasite density. Eight Angolan samples (1.7%) had no or very low PfHRP2 but were positive for 1 or both of the other antigens. PCR analysis confirmed 3 (0.6%) were P. ovale infections and 2 (0.4%) represented P. falciparum parasites lacking Pfhrp2 and/or Pfhrp3.These are the first reports of Pfhrp2/3 deletion mutants in Angola. High-throughput multiplex antigen detection can inexpensively screen for low-density P. falciparum, non-falciparum, and Pfhrp2/3-deleted parasites to provide population-level antigen estimates and identify specimens requiring further molecular characterization.A multiplex malaria antigen detection assay was developed to measure pan-Plasmodium antigens aldolase and LDH, and P. falciparum HRP2. Multiplex antigen detection allowed for prediction of malaria infection status, species of infection, and detection of Pfhrp2/3-deleted parasites.