Search bioRxiv⌕ Search

Biology subjects

Boyce, R. M.

Publications and source records attributed to Boyce, R. M..

2 recordsLinked to original sources

Finding mutations in all the wrong places: Prevalence of knock-down resistance F1534S 1 mutations among Aedes albopictus (family: Culicidae, order: Diptera) in North Carolina

BackgroundKnock-down resistance (kdr) mutations in the voltage gated sodium channel gene of Aedes species mosquitoes are biomarkers for resistance to pyrethroid insecticides. In the United States, few studies have reported kdr mutations among Aedes albopictus (family: Culicidae, order: Diptera, Skuse, 1895) populations. In this study we sought to explore the potential for permethrin-impregnated uniforms worn by military servicemembers to drive kdr emergence. Methods and ResultsWe collected 538 Aedes albopictus mosquitoes, including 156 from 4 sites at Fort Bragg (exposed), North Carolina and 382 from 15 sites in Wake County (control), North Carolina to compare the prevalence of kdr mutations. Of those successfully sequenced, we identified 12 (3.0%) mosquitoes with kdr mutations, all of which were attributed to variants at position 1534 within domain 3. All mutations were found in mosquitoes collected at Wake County sites; no mutations were identified at Fort Bragg. There was a focus of mutations observed at the Wake County sites with approximately 92% (11 of 12) of the mosquitoes with the mutation coming from one site, where kdr mutations represented 24.4% (11 of 45) of all mosquitoes collected. ConclusionsOur study did not show any evidence that universal implementation of permethrin-impregnated uniforms drives the development of resistance. In contrast, we observed highly focal resistance in a suburban area of Raleigh, which may be attributable to peri-domestic mosquito control activities that involve area dispersal of pyrethroid insecticides. More robust surveillance is needed to monitor the emergence and spread of resistance. AUTHOR SUMMARYResistance to commonly employed insecticides among Aedes albopictus (family: Culicidae, order: Diptera, Skuse, 1895) mosquitoes poses as a substantial public health threat. In this study we sought to explore the potential for permethrin-impregnated uniforms worn by military servicemembers to drive emergence of resistance to pyrethroid insecticides by collecting and testing mosquitoes from both military and civilian sites. Overall, we did not identify mosquitoes harboring resistance at Ft. Bragg, but did find a focus of resistance in sub-urban Raleigh, which may be driven by commercial, peri-domestic mosquito control activities. These results suggest that resistance to pyrethroid insecticides may be more prevalent in the United States than previously known, but highly heterogenous. More robust surveillance is needed to monitor the emergence and spread of resistance.

ecology↗

A novel CRISPR-based malaria diagnostic capable of Plasmodium detection, speciation, and drug-resistance genotyping

CRISPR-based diagnostics are a new class of highly sensitive and specific assays with multiple applications in infectious disease diagnosis. SHERLOCK, or Specific High-Sensitivity Enzymatic Reporter UnLOCKing, is one such CRISPR-based diagnostic that combines recombinase polymerase pre-amplification, CRISPR-RNA base-pairing, and LwCas13a activity for nucleic acid detection. We developed SHERLOCK assays for malaria capable of detecting all Plasmodium species known to cause malaria in humans and species-specific detection of P. vivax and P. falciparum, the species responsible for the majority of malaria cases worldwide. We validated these assays against parasite genomic DNA and achieved analytical sensitivities ranging from 2.5-18.8 parasites per reaction. We further tested these assays using a diverse panel of 123 clinical samples from the Democratic Republic of the Congo, Uganda, and Thailand and pools of Anopheles mosquitoes from Thailand. When compared to real-time PCR, the P. falciparum assay achieved 94% sensitivity and 94% specificity in clinical samples. In addition, we developed a SHERLOCK assay capable of detecting the dihydropteroate synthetase (dhps) single nucleotide variant A581G associated with P. falciparum sulfadoxine-pyrimethamine resistance. Compared to amplicon-based deep sequencing, the dhps SHERLOCK assay achieved 73% sensitivity and 100% specificity when applied to a panel of 43 clinical samples, with false-negative calls only at lower parasite densities. These novel SHERLOCK assays have potential to spawn a new generation of molecular diagnostics for malaria and demonstrate the versatility of CRISPR-based diagnostic approaches. One-sentence summaryNovel malaria SHERLOCK assays enabled robust detection, speciation, and genotyping of Plasmodium spp. in diverse samples collected in Africa and Asia.

molecular biology↗