Polymerase Chain Reaction-Based Technique for the Detection of Wuchereria bancrofti in Human Blood Samples, Hydrocele Fluid, and Mosquito Vectors

Siridewa, Kithmini; Karunanayake, E.H.; Chandrasekharan, N.V.

doi:10.4269/ajtmh.1996.54.72

Cited by 28 publications

(10 citation statements)

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“…Como os insetos possuem em seus tecidos, inibidores que podem diminuir a eficiência da reação de PCR, majoritariamente em seu exoesqueleto, cabeça e tórax 13,14 , em nossos experimentos utilizamos dois tipos diferentes de amostras de flebotomíneos para a avaliação dos métodos de extração de DNA. Um lote continha o tubo digestivo do flebotomíneo retirado por centrifugação, e no outro, 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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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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“…The oligonucleotide primers used for the PCR were obtained from (Metabion International AG, Martinsried, Germany). PCR procedure for detection of W. bancrofti in mosquito vectors was executed according to Kithmini et al [ 18 ], using forward pWb12 F (5’-CTGAGTGAAATCAATGAACTGC-3’) and reverse pWb12 R (5’-GTCCATCCGATGAAGTTCCACC-3’) primers. Rishniw et al [ 19 ] described the PCR primers used for detection of D. immitis and D. repens in mosquito vectors, respectively.…”

Section: Methodsmentioning

confidence: 99%

Xenomonitoring of Different Filarial Nematodes Using Single and Multiplex PCR in Mosquitoes from Assiut Governorate, Egypt

et al. 2015

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Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.

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“…The protocol was used to detect parasites in individual mosquitoes and would be for monitoring purposes not cost effective. All recent studies except for one [28] have also used these primers to detect W. bancrofti in mosquitoes. Detection was carried out in pools of 50 Aedes polynesiensis heads, and three DNA extraction protocols were compared for this purpose [6].…”

Section: Detection Of Parasite Dna In Vectorsmentioning

confidence: 99%

Polymerase chain reaction-based detection of lymphatic filariasis

Fischer

Boakye

Hamburger

2003

Med Microbiol Immunol

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PCR-based diagnostic assays are promising tools for the monitoring and evaluation of the Global Programme for Elimination of Lymphatic Filariasis. Sensitive and specific assays have been described for the detection of Wuchereria bancrofti, Brugia malayi, or Brugia timori infection in blood, sputum, and vectors. These techniques can be most cost-effective when employed for pool screening, which is important in the later stages of control programs when infection rates of humans and vectors are low, and large numbers of samples must be examined.

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Polymerase Chain Reaction-Based Technique for the Detection of Wuchereria bancrofti in Human Blood Samples, Hydrocele Fluid, and Mosquito Vectors

Cited by 28 publications

References 0 publications

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

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

Xenomonitoring of Different Filarial Nematodes Using Single and Multiplex PCR in Mosquitoes from Assiut Governorate, Egypt

Polymerase chain reaction-based detection of lymphatic filariasis

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