Brenneman Katelyn Vendrely, Li Xue, Kumar Sudhir, Delgado Elizabeth, Checkley Lisa A, Shoue Douglas A, Reyes Ann, Abatiyow Biley A, Haile Meseret T, Tripura Rupam, Peto Tom, Lek Dysoley, Button-Simons Katrina A, Kappe Stefan H I, Dhorda Mehul, Nosten François, Nkhoma Standwell C, Cheeseman Ian H, Vaughan Ashley M, Ferdig Michael T, Anderson Tim J C
Eck Institute for Global Health, Department of Biological Sciences, University of Notre Dame, Notre Dame, IN, USA.
Program in Disease Intervention and Prevention, Texas Biomedical Research Institute, San Antonio, TX, USA.
iScience. 2022 Mar 16;25(4):104095. doi: 10.1016/j.isci.2022.104095. eCollection 2022 Apr 15.
Classical malaria parasite genetic crosses involve isolation, genotyping, and phenotyping of progeny parasites, which is time consuming and laborious. We tested a rapid alternative approach-bulk segregant analysis (BSA)-that utilizes sequencing of bulk progeny populations with and without drug selection for rapid identification of drug resistance loci. We used dihydroartemisinin (DHA) selection in two genetic crosses and investigated how synchronization, cryopreservation, and the drug selection regimen impacted BSA success. We detected a robust quantitative trait locus (QTL) at in both crosses but did not detect QTLs at four other candidate loci. QTLs were detected using synchronized, but not unsynchronized progeny pools, consistent with the stage-specific action of DHA. We also successfully applied BSA to cryopreserved progeny pools, expanding the utility of this approach. We conclude that BSA provides a powerful approach for investigating the genetic architecture of drug resistance in .
经典的疟原虫遗传杂交涉及子代疟原虫的分离、基因分型和表型分析，这既耗时又费力。我们测试了一种快速替代方法——混合分组分析法（BSA），该方法利用有药物选择和无药物选择的混合子代表型群体的测序来快速鉴定耐药位点。我们在两个遗传杂交中使用双氢青蒿素（DHA）选择，并研究了同步化、冷冻保存和药物选择方案如何影响BSA的成功率。我们在两个杂交中均在 处检测到一个强大的数量性状位点（QTL），但在其他四个候选位点未检测到QTL。使用同步化而非未同步化的子代库检测到了QTL，这与DHA的阶段特异性作用一致。我们还成功地将BSA应用于冷冻保存的子代库，扩展了该方法的实用性。我们得出结论，BSA为研究 中耐药性的遗传结构提供了一种强大的方法。
