A single-round multiplex PCR assay for the identification of Anopheles minimus related species infected with Plasmodium falciparum and Plasmodium vivax

Eamkum, Paiboon; Sungvornyothin, Sungsit; Kritpetcharat, Onanong; Daduang, Jureerut; Lek-Uthai, Usa; Charerntanyarak, Lertchai; Kritpetcharat, Panutas

doi:10.1016/j.parint.2013.11.001

Cited by 6 publications

(8 citation statements)

References 32 publications

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“…Unfortunately, most of these observations relied on species that were identified by morphological characters alone, as molecular assays capable of reliably identifying each sibling species were not readily available or still non‐existent. During the past few decades, studies on malaria vectors in Thailand using molecular identification assays have increased and provided precise identification to the species level, as well as the recognition of additional Anopheles species and species complexes in Thailand (Baimai et al , , Baimai , Green et al , Green et al , Poopittayasataporn and Baimai , Walton et al , Garros et al , Rattanarithikul et al , Dusfour et al , Walton et al , Poolprasert et al , Saeung et al , Thongsahuan et al , Eamkum et al ).…”

Section: Introductionmentioning

confidence: 99%

Diversity of Anopheles species and trophic behavior of putative malaria vectors in two malaria endemic areas of northwestern Thailand

Tainchum

Ritthison

Chuaycharoensuk

et al. 2014

Journal of Vector Ecology

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We determined the species diversity, blood-feeding behavior, and host preference of Anopheles mosquitoes in two malaria endemic areas of Tak (Mae Sot District) and Mae Hong Son (Sop Moei District) Provinces, located along the Thai border with Myanmar, during a consecutive two-year period. Anopheline mosquitoes were collected using indoor and outdoor human-landing captures and outdoor cow-baited collections. Mosquitoes were initially identified using morphological characters, followed by the appropriate multiplex AS-PCR assay for the identification of sibling species within Anopheles (Cellia) complexes and groups present. Real-time PCR was performed for parasite-specific detection in mosquitoes (Plasmodium spp. and Wuchereria bancrofti).

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Section: Introductionmentioning

confidence: 99%

Diversity of Anopheles species and trophic behavior of putative malaria vectors in two malaria endemic areas of northwestern Thailand

Tainchum

Ritthison

Chuaycharoensuk

et al. 2014

Journal of Vector Ecology

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“…Detection of Plasmodium 18S rRNA and An. minimus ITS2A region was performed using the primer set published by Shaw et al [25] and Eamet al [40]. Primer sets are shown in Table 1.…”

Section: Quantitative Amplification Of Wolbachia 16s Rrna and Plasmodmentioning

confidence: 99%

Molecular identification of native Wolbachia pipientis in Anopheles minimus in a low-malaria transmission area of Umphang Valley along the Thailand-Myanmar border

et al. 2020

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Background Wolbachia, obligate intracellular bacteria, infect the majority of arthropods, including many mosquito species of medical importance. Some Wolbachia strains interfere with the development of Plasmodium parasites in female Anopheles, a major vector of malaria. The use of Wolbachia as a means to block malaria transmission is an emerging vector control strategy in highly endemic areas. Hence, identification of native Wolbachia strains in areas where malaria transmission is low may uncover a particular Wolbachia strain capable of Plasmodium interference. This study aims to identify native Wolbachia strains in female Anopheles spp. that are predominant in a low-malaria transmission area in mainland Southeast Asia. Methods Following a 2-year survey of malaria vectors in Umphang Valley of Tak Province, Thailand, DNA extracts of female An. minimus, An. peditaeniatus, and An. maculatus were subjected to amplification of the conserved region of the 16S rRNA-encoding gene. The DNA sequences of the amplicons were phylogenetically compared with those of known Wolbachia strains. Results Among three Anopheles spp., amplification was detected in only the DNA samples from An. minimus. The DNA sequencing of amplicons revealed 100% similarity to Wolbachia pipientis, confirming the specificity of amplification. The Wolbachia-positive An. minimus samples were devoid of Plasmodium 18S rRNA amplification. The phylogenetic trees indicate a close relationship with Wolbachia strains in subgroup B. Conclusion To the best of our knowledge, the data presented herein provide the first molecular evidence of a Wolbachia strain in An. minimus, hereinafter named wAnmi, in a low-malaria transmission area in the Umphang Valley of western Thailand. Further biological characterization is required to examine its potential for malaria transmission control in the field.

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“…In one study, the sensitivities of the ECL-SB and a colorimetric CSP-ELISA for the detection of Pf oocysts in mosquito midguts at 10 days post-infectious feed were determined to be comparable, while the ECL-SB displayed superior detection at earlier time points [20]. RT-PCR has displayed sensitivities of 1.5–75 sporozoites [16, 21]. Our laboratory also recently developed a chemiluminescent ELISA for the detection of P .…”

Section: Introductionmentioning

confidence: 99%

A no film slot blot for the detection of developing P. falciparum oocysts in mosquitoes

et al. 2017

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Non-microscopy-based assays for sensitive and rapid detection of Plasmodium infection in mosquitoes are needed to allow rapid and high throughput measurement of transmission intensity and malaria control program effectiveness. Here, we report on a modified enhanced chemiluminescence-based slot blot assay for detection of Plasmodium falciparum (Pf) circumsporozite protein (PfCSP) expressed on parasite oocysts developing inside the mosquito midgut. This modified assay has several novel features that include eliminating the need for exposure to autoradiography (AR) film, as well as utilizing a novel high affinity anti-CSP antibody, and optimizing assay procedures resulting in significant reduction in the time required to perform the assay. The chemiluminescent signal for the detection of PfCSP in mosquito samples was captured digitally utilizing the C-Digit blot scanner that, allowed the detection of 0.01 pg of recombinant P. falciparum CSP and as few as 0.02 P. falciparum oocysts in a little over two hours. The earlier ECL-SB detected rCSP and oocysts and took approximately 5 h to perform. Whole mosquito lysates from both high and low prevalence—infected mosquito populations were prepared and evaluated for PfCSP detection on the ECL-SB by both AR film and digital data capture and analysis. There was a 100% agreement between the AR film and the C-Digit scanner methods for PfCSP detection in randomly sampled mosquitoes. This novel “No Film” Slot Blot assay obviates the need for AR film exposure and development and significantly reduces the assay time enabling widespread use in field settings.

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A single-round multiplex PCR assay for the identification of Anopheles minimus related species infected with Plasmodium falciparum and Plasmodium vivax

Cited by 6 publications

References 32 publications

Diversity of Anopheles species and trophic behavior of putative malaria vectors in two malaria endemic areas of northwestern Thailand

Diversity of Anopheles species and trophic behavior of putative malaria vectors in two malaria endemic areas of northwestern Thailand

Molecular identification of native Wolbachia pipientis in Anopheles minimus in a low-malaria transmission area of Umphang Valley along the Thailand-Myanmar border

A no film slot blot for the detection of developing P. falciparum oocysts in mosquitoes

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