Genetic screen for P. vivax CQR

描述（由申请人提供）：对氯喹耐药的间日疟原虫（CQ）被广泛报道，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。由于间日疟原虫氯喹耐药（chloroquine resistance， CQR）决定因子和其他关键分子的同源物未发现与间日疟原虫CQR有关，因此间日疟原虫产生CQR的机制显然与恶性疟原虫不同。因此，我们建议采用全基因组重组的方法，将CQR型间日疟原虫全基因组cDNA克隆到piggyBac载体中，转染到氯喹敏感（CQS）培养的恶性疟原虫中。转染后的恶性疟原虫将在CQ压力下培养，并通过限制稀释进行克隆。含cdna的质粒将从生长中的寄生虫克隆中提取出来，以鉴定间日疟原虫CQR中起重要作用的候选基因。该结果将能够对CQR间日疟原虫的传播进行大规模监测和监测，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. PUBLIC HEALTH RELEVANCE: Plasmodium vivax is the major cause of malaria outside of Africa and is has developing resistance to chloroquine (CQR), which is the first line treatment, thereby impeding effective case management and control. Genetic traits underlying P. vivax CQR are different from P. falciparum and cannot be identified by a simple 'copy cat' approach. We propose a novel whole genome genetic screening approach to identify P. vivax CQR resistance genes by transgene complementation and phenotype selection of P falciparum.