Genetic screen for P. vivax CQR

描述（由申请方提供）：间日疟原虫对氯喹（CQ）耐药的报道广泛，严重阻碍了病例管理和疟疾控制/消除工作。耐药的分子决定因素和机制尚不清楚。由于已发现恶性疟原虫氯喹抗性（CQR）决定簇和其他关键分子的同源物与间日疟原虫CQR无关，因此很明显间日疟原虫使用与恶性疟原虫不同的机制发展了CQR。因此，我们提出使用全基因组重组工程类型的方法，由此将CQR间日疟原虫基因组的整个cDNA克隆到piggyBac载体中并转染到恶性疟原虫的氯喹敏感（CQS）培养系中。将在CQ压力下培养转染的恶性疟原虫寄生虫，并通过有限稀释进行克隆。将从具有生长的寄生虫的克隆中拯救含有cDNA的质粒，以鉴定在间日疟原虫CQR中起重要作用的候选基因。该结果将能够大规模监测和监测CQR间日疟原虫的传播，为治疗结果提供预测价值，并为政策变化提供信息。

DESCRIPTION (provided by applicant): Plasmodium vivax resistant to chloroquine (CQ) is widely reported, seriously hampering the case management and malaria control/elimination effort. The molecular determinant and mechanism of resistance is unclear. As homologs of P. falciparum chloroquine resistance (CQR) determinant and other key molecules have been found not implicated in P. vivax CQR, it is clear that P. vivax has developed CQR using a mechanism different to P. falciparum. Therefore, we propose to use a whole genome recombineering type of approach, whereby the entire cDNA of a CQR P. vivax genome is cloned into piggyBac vector and transfected into a chloroquine sensitive (CQS) cultured line of P. falciparum. The transfected P. falciparum parasites will be cultured under CQ pressure and cloned by limiting dilution. cDNA-containing plasmids will be rescued from clones with growing parasites to identify candidate gene(s) that plays significant roles in P. vivax CQR. The outcome will enable large-scale monitor and surveillance of the spread of CQR P. vivax, provide predictive value for treatment outcome and provide information for policy changes.