Detection of &lt;em&gt;Plasmodium&lt;/em&gt; Sporozoites in &lt;em&gt;Anopheles&lt;/em&gt; Mosquitoes using an Enzyme-linked Immunosorbent Assay

Kumpitak, Chalermpon; Nguitragool, Wang; Sattabongkot, Jetsumon; Bantuchai, Sirasate

doi:10.3791/63158

Cited by 5 publications

(6 citation statements)

References 18 publications

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“…Historically, SGs have been regarded as a difficult tissue to dissect en masse, thus limiting the scope of analysis. Following various refinements in adult mosquito SG studies 2 , 3 , 4 , this research group developed a powerful means to view the adult tissue's larval precursor 1 6 . This method has distinct advantages when compared to microscopy, and it can be applied broadly across parasite subtypes.…”

Section: Chiu Et Al Provide a Clear Detailed Explanation Of Howmentioning

confidence: 99%

Highlighting Molecular Biology as a Focal Point for Mosquito Research

Wells¹

2023

JoVE

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Section: Chiu Et Al Provide a Clear Detailed Explanation Of Howmentioning

confidence: 99%

Highlighting Molecular Biology as a Focal Point for Mosquito Research

Wells¹

2023

JoVE

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“…Malaria is a vector-borne disease transmitted by Anopheles mosquitoes [7][8][9]. The WHO recorded 4500 species of mosquitoes worldwide in 34 genera from the family Culicidae, the order Diptera, the class Insecta, and the phylum Arthropoda [10].…”

Section: Introductionmentioning

confidence: 99%

Updating the Data on Malaria Vectors in Malaysia: Protocol for a Scoping Review

Ali¹,

Ali²,

Putit³

2023

JMIR Res Protoc

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Background Malaria is still a public health threat. From 2015 to 2021, a total of 23,214 malaria cases were recorded in Malaysia. Thus, effective intervention and key entomological information are vital for interrupting or preventing malaria transmission. Therefore, the availability of malaria vector information is desperately needed. Objective The objective of our study is to update the list of human and zoonotic malaria vectors in Malaysia. This work will include (1) the characterization of the key behavioral traits and breeding sites of malaria vectors and (2) the determination of new and potential malaria vectors in Malaysia. The findings of our scoping review will serve as decision-making evidence that stakeholders and decision makers can use to strengthen and intensify malaria surveillance in Malaysia. Methods The scoping review will be conducted based on the following four electronic databases: Scopus, PubMed, Google Scholar, and Science Direct. A search strategy was conducted for articles published from database inception to March 2022. The criteria for article inclusion were any malaria vector–related studies conducted in Malaysia (with no time frame restrictions) and peer-reviewed studies. The PRISMA-ScR (Preferred Reporting Items for Systematic Reviews and Meta-Analyses Extension for Scoping Reviews) will be used to guide our systematic approach. Data from published research literature will be extracted by using a standardized data extraction framework, including the titles, abstracts, characteristics, and main findings of the included studies. To assess the risk of bias, articles will be screened independently by 2 reviewers, and a third reviewer will make the final decision if disagreements occur. Results The study commenced in June 2021, and it is planned to be completed at end of 2022. As of early 2022, we identified 631 articles. After accessing and evaluating the articles, 48 were found to be eligible. Full-text screening will be conducted in mid-2022. The results of the scoping review will be published as an open-access article in a peer-reviewed journal. Conclusions Our novel scoping review of malaria vectors in Malaysia will provide a comprehensive evidence summary of updated, relevant information. An understanding of the status of Anopheles as malaria vectors and the knowledge generated from the behavioral characteristics of malaria vectors are the key components in making effective interventions for eliminating malaria. International Registered Report Identifier (IRRID) DERR1-10.2196/39798

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“…To overcome the limitations of microscopy a number of assays have been developed to enable high throughput detection of parasites in the mosquito gut and salivary glands. These assays include ELISA to detect the circumsporozoite protein (CSP) in mosquito lysates (CSP-ELISA) (Beier et al, 1987;Kumpitak et al, 2021;Sutcliffe et al, 2021), bioluminescence assays to detect transgenic parasites with the green fluorescence protein (GFP) (Delves and Sinden, 2010;Stone et al, 2014;Singer and Frischknecht, 2021), near-infrared spectroscopy (NIRS) to detect parasites within mosquitoes (Maia et al, 2019;Da et al, 2021), enhanced chemiluminescent slot blot (ECL-SB) for detecting PfCSP in mosquito samples (Kumar et al, 2014;Grabias et al, 2017) and molecular detection of Plasmodium DNA (Boissière et al, 2013;Marie et al, 2013;Sazed et al, 2021). Although the CSP-ELISA is relatively robust and cost effective it is only semi quantitative (Beier et al, 1987;Kumpitak et al, 2021;Sutcliffe et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

Using qPCR to compare the detection of Plasmodium vivax oocysts and sporozoites in Anopheles farauti mosquitoes between two DNA extraction methods

Timinao

Jamea²,

Katusele³

et al. 2023

Front. Parasitol.

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BackgroundCurrently, the gold standard to assess parasite developmental stages in mosquitoes is light microscopy. Microscopy can miss low-density infections, is time-consuming and not species-specific. Enzyme-linked immunosorbent assay (ELISA) has been the alternative technique to evaluate the infectivity of mosquitoes especially in field studies however it is semi-quantitative. Molecular techniques that have been used to detect the mosquito stages of malaria parasites including P. vivax. Here, we present a quantitative real-time assay (qPCR) that can be used to detect low-density P. vivax oocyst and sporozoite infections while comparing parasites extracted by the conventional DNA extraction and heating methods.MethodsColony reared Anopheles farauti mosquitoes were exposed to blood samples collected from infected individuals using a direct membrane feeding assay. The fully fed mosquitoes were kept for 7 and 14 days post-feed before dissection to confirm presence of oocysts and sporozoites. Infected mosquito guts and the salivary glands (with the head and thorax) were stored and DNA was extracted either by heating or by performing conventional column-based DNA extraction. Following DNA extraction the infected samples were subjected to qPCR to detect P. vivax parasites.ResultsDNA extraction of 1 or more oocysts by heating resulted in an overall sensitivity of 78% (57/73) and single oocysts infections were detected with a sensitivity of 82% (15/17) in the heating arm. We observed a 60% (18/30) sensitivity with sporozoites where DNA was extracted using the conventional DNA extraction method. We show that the heating method significantly improved the detection of oocysts over conventional DNA extraction. There was no significant difference in the DNA copy numbers when comparing the detection of oocysts from the conventional DNA extraction versus heating. However, we observed that the DNA copy numbers of the sporozoites detected in the heating arm was significantly higher than in the conventional DNA extraction arm.ConclusionWe have adapted a qPCR assay which, when coupled with heating to release DNA reduces sample processing time and cost. Direct qPCR after heating will be a useful tool when investigating transmission blocking vaccines or antimalarials or when evaluating field caught mosquitoes for the presence of malaria parasites.

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Detection of <em>Plasmodium</em> Sporozoites in <em>Anopheles</em> Mosquitoes using an Enzyme-linked Immunosorbent Assay

Cited by 5 publications

References 18 publications

Highlighting Molecular Biology as a Focal Point for Mosquito Research

Highlighting Molecular Biology as a Focal Point for Mosquito Research

Updating the Data on Malaria Vectors in Malaysia: Protocol for a Scoping Review

Using qPCR to compare the detection of Plasmodium vivax oocysts and sporozoites in Anopheles farauti mosquitoes between two DNA extraction methods

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