HRP2 and HRP3 cross-reactivity and implications for HRP2-based RDT use in regions with Plasmodium falciparum hrp2 gene deletions.
HRP2 and HRP3 cross-reactivity and implications for HRP2-based RDT use in regions with Plasmodium falciparum hrp2 gene deletions.
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DOI：
10.1186/s12936-021-03739-6
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发表时间：
2021-04-29
期刊：
影响因子：
3
通讯作者：
Aidoo M
中科院分区：
文献类型：
--
作者：
Kong A;Wilson SA;Ah Y;Nace D;Rogier E;Aidoo M
恶性疟原虫富含组氨酸的抗原蛋白2（HRP 2）是疟疾快速诊断试验（RDTs）的首选靶点，因为其大量产生的寄生虫和热稳定性。因此，全球采购的大多数RDT都靶向这种抗原。然而，来自南美洲的先前报告和来自撒哈拉以南非洲和亚洲的最近报告表明某些恶性疟原虫寄生虫具有编码HRP 2的基因的缺失。HRP 2抗原与另一种恶性疟原虫抗原HRP 3是旁系同源的，并且HRP 2的一些抗体与HRP 3交叉反应。已经描述了多种具有hrp 2和hrp 3基因之一或两者缺失的寄生虫。目前还不清楚hrp 2和hrp 3缺失基因型的各种组合如何影响基于HRP 2的RDT的临床敏感性。在疟疾RDT上使用培养适应的恶性疟原虫寄生虫测试HRP 2和HRP 3之间的交叉反应性，所述恶性疟原虫寄生虫具有hrp 2和hrp 3完整或具有一个或两个基因缺失。制备4种培养适应性恶性疟原虫寄生虫[3D 7（hrp 2 +/hrp 3+）、Dd 2（hrp 2 −/hrp 3+）、HB 3（hrp 2 +/hrp 3 −）和3BD 5（hrp 2 −/hrp 3 −）]的10倍系列稀释液，范围为100，000至0.01寄生虫/µL。使用多重微珠测定法测定稀释样品的HRP 2、疟原虫乳酸脱氢酶（pLDH）和醛缩酶浓度。随后对三种RDT产品进行了检测，这些产品旨在通过HRP 2单独或与pLDH组合检测恶性疟原虫。在寄生虫密度约为1000个寄生虫/µL时，所有三种RDT均检出表达hrp 2或hrp 3的寄生虫。与缺失一个基因的HB 3（3.02 ng/mL）和Dd 2（0.20 ng/mL）菌株相比，当hrp 2和hrp 3基因均完整时（3D 7寄生虫，47.9 ng/mL），使用HRP 2偶联微珠进行的基于多重抗原的测定显示出更高的抗原浓度。10个寄生虫/µL（0.45 ng/mL）的3D 7对所有三种RDT产品均具有反应性，而在该密度下，其他寄生虫均无反应性。在一定的抗原阈值以上，HRP 3对基于HRP 2的RDT的交叉反应性足以掩盖仅hrp 2缺失的影响。hrp 2缺失及其对基于hrp 2的RDT的影响的研究必须与hrp 3缺失一起研究，并包括基于hrp 2的检测的临床样本反应性。
The Plasmodium falciparum antigen histidine rich protein 2 (HRP2) is a preferred target for malaria rapid diagnostic tests (RDTs) because of its abundant production by the parasite and thermal stability. As a result, a majority of RDTs procured globally target this antigen. However, previous reports from South America and recent reports from sub-Saharan Africa and Asia indicate that certain P. falciparum parasites have deletions of the gene coding for HRP2. The HRP2 antigen is paralogous to another P. falciparum antigen HRP3 and some antibodies to HRP2 cross-react with HRP3. Multiple parasites have been described with deletions of one or both hrp2 and hrp3 genes. It is unclear how the various combinations of hrp2 and hrp3 deletion genotypes affect clinical sensitivity of HRP2-based RDTs. Cross-reactivity between HRP2 and HRP3 was tested on malaria RDTs using culture-adapted P. falciparum parasites with both hrp2 and hrp3 intact or with one or both genes deleted. Ten-fold serial dilutions of four culture-adapted P. falciparum parasites [3D7 (hrp2+/hrp3+), Dd2 (hrp2−/hrp3+), HB3 (hrp2+/hrp3−) and 3BD5 (hrp2−/hrp3−)] ranging from 100,000 to 0.01 parasites/µL were prepared. HRP2, Plasmodium lactate dehydrogenase (pLDH) and aldolase concentrations were determined for the diluted samples using a multiplex bead assay. The samples were subsequently tested on three RDT products designed to detect P. falciparum by HRP2 alone or in combination with pLDH. At parasite densities of approximately 1000 parasites/µL, parasites that expressed either hrp2 or hrp3 were detected by all three RDTs. Multiplex based antigen measurement using HRP2- conjugated beads demonstrated higher antigen concentration when both hrp2 and hrp3 genes were intact (3D7 parasites, 47.9 ng/ml) compared to HB3 (3.02 ng/mL) and Dd2 (0.20 ng/mL) strains that had one gene deleted. 3D7 at 10 parasites/µL (0.45 ng/mL) was reactive on all three RDT products whereas none of the other parasites were reactive at that density. Above a certain antigen threshold, HRP3 cross-reactivity on HRP2-based RDTs is sufficient to mask the effects of deletions of hrp2 only. Studies of hrp2 deletion and its effects on HRP2-based RDTs must be studied alongside hrp3 deletions and include clinical sample reactivity on HRP2-based tests.
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