Plasmodium sp.: Optimal Protocols for PCR Detection of Low Parasite Numbers from Mosquito (Anopheles sp.) Samples

Arez, Ana Paula; Lopes, Dinora; Pinto, João; Franco, Ana Sofia; Snounou, Georges; Rosário, Virgílio Estólio do

doi:10.1006/expr.2000.4496

Cited by 45 publications

(36 citation statements)

References 16 publications

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“…Therefore, lower infection rates in the vectors in Minami Daito Island (1.2%) might associate with their origin compared to the artificially introduced vector in Hawaii (15%). Detection sensitivity must be strongly considered because recent remarkable progress of ELISA and/or PCR methods enables more feasible results [2,9,37]. In the present study, annual prevalence of positive rate in vectors still remains unclear.…”

Section: Discussionmentioning

confidence: 78%

“…Direct detection of infective stage protozoa from their vectors is accompanied with difficulties in the requirement for trained skills and in the case of false positive because prevalence of pathogens in nature tends to be relatively low levels [15]. Although several studies showed advantages of molecular detection of various Plasmodium genes from mosquitoes using PCR [2,6,21,36,37], no molecular based investigations have been reported on the detection of avian malarial protozoa from wild mosquitoes of Japan, neither even on morphological detection of sporozoites. In this study, we detected avian Plasmodium DNA sequences from the wild mosquitoes collected in Japan for the first time.…”

Section: Discussionmentioning

confidence: 99%

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Detection of Avian Plasmodium spp. DNA Sequences from Mosquitoes Captured in Minami Daito Island of Japan

Ejiri

Sato

Sasaki

et al. 2008

The Journal of Veterinary Medical Science

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ABSTRACT. Several species of birds in Minami Daito Island, an oceanic island located in the far south from the main islands of Japan, were found to be infected with avian Plasmodium. However, no vector species of the avian malaria in this island have been revealed yet. To speculate potential vectors, we collected mosquitoes there and investigated using a PCR procedure whether the mosquitoes harbor avian malaria or not. Totally 1,264 mosquitoes including 9 species were collected during March 2006 to February 2007. The mosquitoes collected were stored every species, sampled date and location for DNA extraction. Fifteen out of 399 DNA samples showed positive for the partial mtDNA cytb gene of avian Plasmodium. Estimated minimum infection rate among collected mosquitoes was 1.2% in this study. Four species of mosquitoes; Aedes albopictus, Culex quinquefasciatus, Lutzia fuscanus and Mansonia sp. had avian Plasmodium gene sequences. Detected DNA sequences from A. albopictus and L. fuscanus were identical to an avian Plasmodium lineage detected in bull-headed shrike (Lanius bucephalus) captured in the island. Different sequences were detected from C. quinquefasciatus, which were corresponding to an avian Plasmodium from a sparrow (Passer montanus) and Plasmodium gallinaceum. Our results suggest that A. albopictus, Lutzia fuscanus, C. quinquefasciatus, and Mansonia sp. could be potential vectors of avian malaria in Minami Daito Island. This study was the first report of molecular detection of avian Plasmodium from mosquitoes in Japan.

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Section: Discussionmentioning

confidence: 78%

Section: Discussionmentioning

confidence: 99%

Detection of Avian Plasmodium spp. DNA Sequences from Mosquitoes Captured in Minami Daito Island of Japan

Ejiri

Sato

Sasaki

et al. 2008

The Journal of Veterinary Medical Science

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“…Resultados similares foram obtidos por Arez et al 16 em seus experimentos com identificação de Plasmodium em mosquitos do gênero Anopheles dissecados (conteúdo intestinal, cabeça e tórax) ou inteiros. Esses autores verificaram inibição da reação de PCR pela presença dos inibidores nos tecidos dos insetos, e esta foi particularmente marcada quando se utilizou o protocolo que inclui passagens de fenol/clorofórmio/ál-cool isoamílico em insetos inteiros.…”

Section: Discussionunclassified

Padronização de condições para detecção de DNA de Leishmania spp. em flebotomíneos (Diptera, Psychodidae) pela reação em cadeia da polimerase

Paiva¹,

Secundino

Pimenta

et al. 2007

Cad. Saúde Pública

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A correta identificação dos agentes etiológicos em insetos vetores é de crucial importância aos estudos epidemiológicos. A pesquisa de flagelado nesses vetores, pela dissecção de seu trato digestivo, observação microscópica do seu conteúdo ou por isolamento dos parasitas provenientes de insetos em meios de cultura, tem-se mostrado operacionalmente inadequada e com baixa especificidade do diagnóstico, pois fêmeas de flebotomíneos também podem albergar outros flagelados como Trypanosoma e Endotrypanum. Acreditamos que por sua eficiência e especificidade, a amplificação de seqüências-alvo do DNA da Leishmania, por meio da reação em cadeia de polimerase, pode ser aplicada na investigação de sua presença em flebotomíneos, desde que estes estejam devidamente acondicionados e o DNA do parasita extraído a partir de metodologia adequada. Este trabalho descreve metodologias utilizadas na padronização da conservação dos espécimes de flebotomíneos e extração do DNA da Leishmania como uma alternativa mais prática que os métodos tradicionais.

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“…Because sporozoites, the infective stage of parasites, are located in the salivary glands, genomic DNA from the head and thorax of mosquitoes was processed for molecular analysis. Because loss of polymerase chain reaction (PCR) amplification efficiency is likely to result from inhibitors present in the mosquito tissues, 17 we used the IsoQuick DNA isolation kit (ORCA Research, Bothell, WA), a silica/guanidinium-based template preparation method, 18 that efficiently remove PCR inhibitors 17 on parasite detection from infected mosquitoes.…”

Section: Studymentioning

confidence: 99%

Monitoring Chloroquine Resistance Using Plasmodium Falciparum Parasites Isolated From Wild Mosquitoes in Tanzania

Temu

Kimani²,

Tuno³

et al. 2006

The American Journal of Tropical Medicine and Hygiene

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Abstract. Monitoring antimalarial drug resistance is a useful epidemiologic tool and provides early detection of resistance foci. Using DNA extracted from the head/thorax of wild mosquitoes collected from Bagamoyo Coastal Tanzania, samples infected by Plasmodium falciparum (N ‫ס‬ 89, in 2002 and N ‫ס‬ 249 in 2004) were screened by nested polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assay for mutations at Pfcrt76 and Pfmdr1-86 associated with chloroquine (CQ) resistance. The majority of isolates were of single infection (71%), and the prevalence of mutant alleles of Pfcrt76 decreased from 64.5% in 2002 to 16% in 2004; likewise, mutant Pfmdr1-86 alleles decreased from 46.6% to 2.7%. Overall, there was a decline of mutant isolates by a factor of 17 and 4 for Pfmdr1 and Pfcrt, respectively. In contrast, isolates with wild-type alleles increased significantly from < 20% in 2002 to 67.6% for Pfcrt76 and 83.5% for Pfmdr1-86 in 2004. This observation suggest a biologic trend of decrease of CQ mutants and a subsequent increase of CQ susceptible parasites in circulation after the discontinued use of CQ in 2001 as a first-line drug in Tanzania. High prevalence of susceptible P. falciparum found in circulation not only supports other reports of a decline of mutant parasites after a reduction of drug selection pressure but suggests that the fitness cost is high in mutant parasites. Typing parasite isolates from infected mosquitoes, an alternative means of data collection, has the potential to increase the spatial and temporal coverage, and this approach is practical in highly endemic regions of Africa.

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Plasmodium sp.: Optimal Protocols for PCR Detection of Low Parasite Numbers from Mosquito (Anopheles sp.) Samples

Cited by 45 publications

References 16 publications

Detection of Avian Plasmodium spp. DNA Sequences from Mosquitoes Captured in Minami Daito Island of Japan

Detection of Avian Plasmodium spp. DNA Sequences from Mosquitoes Captured in Minami Daito Island of Japan

Padronização de condições para detecção de DNA de Leishmania spp. em flebotomíneos (Diptera, Psychodidae) pela reação em cadeia da polimerase

Monitoring Chloroquine Resistance Using Plasmodium Falciparum Parasites Isolated From Wild Mosquitoes in Tanzania

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