A Multi-detection Assay for Malaria Transmitting Mosquitoes

Y, Lee; Weakley, Allison M.; Nieman, Catelyn C.; Malvick, Julia; Lanzaro, Gregory C.

doi:10.3791/52385

Cited by 19 publications

(17 citation statements)

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“…The iPLEX method allows the multiplex detection of up to 40 SNPs in a single reaction and can be completed within 5 hours after gDNA extraction. The economical high multiplexing capacity of iPLEX assays provides increased diagnostic accuracy when compared with other PCR-based techniques (Lee et al 2015).…”

Section: Discussionmentioning

confidence: 99%

“…However, using a combination of the SNP markers will provide higher confidence for species identification by reducing false positives (Lee et al 2015), given the presence of genetic variations in field populations. Other genotyping technologies mentioned above can be used in combination with the SNP markers generated in this study for T. absoluta species diagnostics, but the multiplexing capacity of some of these technologies, e.g.…”

Section: Discussionmentioning

confidence: 99%

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Sequencing of Tuta absoluta genome to develop SNP genotyping assays for species identification

et al. 2019

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Tuta absoluta is one of the most devastating pests of fresh market and processing tomatoes. Native to South America, its detection was confined to that continent until 2006 when it was identified in Spain. It has now spread to almost every continent, threatening countries whose economies rely heavily on tomatoes. This insect causes damage to all developmental stages of its host plant, leading to crop losses as high as 80 to 100%. Although T. absoluta has yet to be found in the U.S. and China, which makes up a large portion of the tomato production in the world, computer models project a high likelihood of invasion. To halt the continued spread of T. absoluta and limit economic loss associated with tomato supply chain, it is necessary to develop accurate and efficient methods to identify T. absoluta and strengthen surveillance programs. Current identification of T. absoluta relies on examination of morphology and assessment of host plant damage, which are difficult to differentiate from that of native tomato pests. To address this need, we sequenced the genomes of T. absoluta and two closely related Gelechiidae, Keiferia lycopersicella, and Phthorimaea operculella, and developed a bioinformatic pipeline to design a panel of 21 SNP markers for species identification. The accuracy of the SNP panel was validated in a multiplex format using the iPLEX chemistry of Agena MassARRAY system. Finally, the new T. absoluta genomic resources we generated can be leveraged to study T. absoluta biology and develop species-specific management strategies. Key Message  Surveillance and identification of Tuta absoluta are challenging because it is morphologically similar to closely related species, e.g. Keiferia lycopersicella and Phthorimaea operculella.  We generated new genomic sequences for these three species and identified single nucleotide polymorphisms (SNPs) to facilitate species identification.  We validated a multiplex genotyping panel of 21 SNPs using the iPLEX MassARRAY platform and confirmed its accuracy for species identification.  We generated new molecular tools and genome resources to aid Tuta absoluta management. leaves, where the larvae will emerge from eggs and begin mining host tissues. Larvae can also enter the stems through the buds, and feed within the tomato fruit, leaving them unmarketable. Its distribution was largely confined to South America until it was first detected in Spain in 2006 (Desneux et al. 2010; Guillemaud et al. 2015). Since then T. absoluta has spread rapidly and is now

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Sequencing of Tuta absoluta genome to develop SNP genotyping assays for species identification

et al. 2019

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“…The specific host species that each mosquito had fed upon was determined by a multiplex genotyping assay on DNA extracted from abdomens (Lee et al 2015). This multiplex genotyping assay can distinguish between blood from cattle, goat, pig, dog, chicken and human.…”

Section: Bloodmeal Analysismentioning

confidence: 99%

The genetic basis of host choice and resting behavior in the major African malaria vector, Anopheles arabiensis

Main

Ferguson

et al. 2016

Preprint

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Malaria transmission is dependent on the propensity of Anopheles mosquitoes to bitehumans (anthropophily) instead of other dead end hosts. Recent increases in the usage of Long Lasting Insecticide Treated Nets (LLINs) in Africa have been associated with reductions in highly anthropophilic vectors such as Anopheles gambiae s.s.,leaving less anthropophilic species such as Anopheles arabiensis as the most prominent remaining source of transmission in many settings. An.arabiensis is more of a generalist in terms of its host choice and resting behavior, which may be due to phenotypic plasticity and/or segregating allelic variation. To investigate the potential genetic basis of host choice and resting behavior in An. arabiensis we performed a genome-wide association study on host choice (human-or cattle-fed) and resting position (collected indoors or outdoors) in the Kilombero Valley, Tanzania. This represents the first genomic/molecular analysis of host choice and resting behavior in a malaria vector. We identified a total of 4,820,851 SNPs, which were used to conduct the first genome-wide estimates of 'SNP heritability' for host choice and resting behavior in this species. A genetic component was detected for host choice (human vs cow fed; permuted P = 0.002), but there was no evidence of a genetic component for resting behavior (indoors versus outside; permuted P = 0.465). A principal component analysis (PCA) segregated individuals based on genomic variation into three groups which are characterized by differences at the 2Rb and/or 3Ra paracentromeric chromosome inversions. There was a non-random distribution of cattle-fed mosquitoes between the PCA clusters, suggesting that alleles linked to the 2Rb and/or 3Ra inversions may influence host choice. Using a novel inversion genotyping assay, we detected a significant enrichment of the standard arrangement (non-inverted) of 3Ra among cattle-fed mosquitoes (N=129) versus all non-cattle-fed individuals (N=234; १2, p=0.007). Thus, tracking the frequency of the 3Ra in An. arabiensis populations is important, especially in relation to the emergence of behavioral avoidance(e.g. shifting toward cattle-feeding) in some populations. A better understanding of the genetic basis for host choice in An. arabiensis may also open avenues for novel vector control strategies based on introducing genes for zoophily into wild mosquito populations.Author summaryMalaria transmission is driven by the propensity for mosquito vectors to bite people, whilst its control depends on the tendency of mosquitoes to bite and rest in places where they will come into contact with insecticides. In many parts of Africa, Anopheles arabiensis is now the only remaining vec 63 tor in areas where coverage with Long Lasting Insecticide Treated Nets is high. We sought to assess the potential for An. arabiensis to adapt its behavior to avoid control measures by investigating the genetic basis for its host choice and resting behavior. Blood fed An. arabiensis were collected resting indoors and outdoors in the Kilombero Valley, Tanzania. We sequenced a total of 48 genomes representing 4 phenotypes (human or cow fed, resting in or outdoors) and tested for a genetic basis for each phenotype. Genomic analysis followed up by application of a novel molecular karyotyping assay revealed a relationship between An. arabiensis that fed on cattle and the standard arrangement of the 3Ra inversion. This indicates that the host choice behavior of An. arabiensis has has a substantial genetic component. Validation with controlled host preference assays comparing individuals with the standard and inverted arrangement of 3Ra is still needed.

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“…Additionally, there is concern that progress may have stalled after more than a decade of success in global malaria control 2 . Of major concern is the increase in insecticide resistance among mosquito populations throughout Africa 3 , which is degrading the lethality and effectiveness of vector control tools, notably indoor residual spraying (IRS) and long-lasting insecticide treated nests (LLINs) which have been the cornerstones of malaria control in the past decades 4 . Indeed, much of the effectiveness of LLINs and IRS comes from community-wide reductions in vector population size, not merely from preventing people from getting bitten 5 .…”

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confidence: 99%

Prediction of mosquito species and population age structure using mid-infrared spectroscopy and supervised machine learning

González‐Jiménez¹,

Babayan²,

Khazaeli³

et al. 2019

Wellcome Open Res

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Despite the global efforts made in the fight against malaria, the disease is resurging. One of the main causes is the resistance that Anopheles mosquitoes, vectors of the disease, have developed to insecticides. must survive for at least 10 days to possibly transmit malaria. Anopheles Therefore, to evaluate and improve malaria vector control interventions, it is imperative to monitor and accurately estimate the age distribution of mosquito populations as well as their population sizes. Here, we demonstrate a machine-learning based approach that uses mid-infrared spectra of mosquitoes to characterise simultaneously both age and species identity of females of the African malaria vector species Anopheles and . mid-infrared spectroscopy-based prediction of gambiae An. arabiensis mosquito age structures was statistically indistinguishable from true modelled distributions. The accuracy of classifying mosquitoes by species was 82.6%. The method has a negligible cost per mosquito, does not require highly trained personnel, is rapid, and so can be easily applied in both laboratory and field settings. Our results indicate this method is a promising alternative to current mosquito species and age-grading approaches, with further improvements to accuracy and expansion for use Any reports and responses or comments on the article can be found at the end of the article. approaches, with further improvements to accuracy and expansion for use with other mosquito vectors possible through collection of larger mid-infrared spectroscopy data sets.

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A Multi-detection Assay for Malaria Transmitting Mosquitoes

Cited by 19 publications

References 40 publications

Sequencing of Tuta absoluta genome to develop SNP genotyping assays for species identification

Sequencing of Tuta absoluta genome to develop SNP genotyping assays for species identification

The genetic basis of host choice and resting behavior in the major African malaria vector, Anopheles arabiensis

Prediction of mosquito species and population age structure using mid-infrared spectroscopy and supervised machine learning

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