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Xenomonitoring of different filarial nematodes using single and multiplex PCR in mosquitoes from Assiut Governorate, Egypt.

Dyab AK, Galal LA, Mahmoud Ael-S, Mokhtar Y - Korean J. Parasitol. (2015)

Bottom Line: The technique showed 100% sensitivity and 98% specificity.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Parasitology, Faculty of Medicine, Assiut University, Assiut, Egypt.

ABSTRACT
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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Multiplex PCR pattern on 2% ethidium bromide-stained agarose gel. Lane 1, W. bancrofti; lane 2, D. immitis; lane 3, D. repens. (A) Multiplex PCR confirmed the results of single PCR in El-Nikhila, El-Matiaa villages and Sahel Seleem district. (B) Multiplex PCR detected a positive sample of W. bancrofti (lane 1) that could not be detected by single PCR in El-Nikhila village. (C) Distribution of filarial parasites among different mosquito genera using multiplex PCR. Lane 1, Culex spp.; lane 2, Anopheles spp.; lane 3, Aedes spp. (D) Distribution of filarial parasites among different Culex species in El-Nikhila village. Lane 1, Culex pipiens; lane 2, Culex antennatus; lane 3, Culex pusillus.
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f3-kjp-53-1-77: Multiplex PCR pattern on 2% ethidium bromide-stained agarose gel. Lane 1, W. bancrofti; lane 2, D. immitis; lane 3, D. repens. (A) Multiplex PCR confirmed the results of single PCR in El-Nikhila, El-Matiaa villages and Sahel Seleem district. (B) Multiplex PCR detected a positive sample of W. bancrofti (lane 1) that could not be detected by single PCR in El-Nikhila village. (C) Distribution of filarial parasites among different mosquito genera using multiplex PCR. Lane 1, Culex spp.; lane 2, Anopheles spp.; lane 3, Aedes spp. (D) Distribution of filarial parasites among different Culex species in El-Nikhila village. Lane 1, Culex pipiens; lane 2, Culex antennatus; lane 3, Culex pusillus.

Mentions: Afterwards, multiplex PCR had confirmed the single PCR results performed in the 3 localities (Fig. 3A), in fact multiplex PCR was superior to single PCR as it positively detected W. bancrofti in previously negative tested pool (Fig. 3B). The sensitivity of multiplex PCR as compared to single PCR was 100%, and the specificity was 98%. The positive predictive value was 93%, the negative predictive value was 100%, and the accuracy rate was 99% (Table 1).


Xenomonitoring of different filarial nematodes using single and multiplex PCR in mosquitoes from Assiut Governorate, Egypt.

Dyab AK, Galal LA, Mahmoud Ael-S, Mokhtar Y - Korean J. Parasitol. (2015)

Multiplex PCR pattern on 2% ethidium bromide-stained agarose gel. Lane 1, W. bancrofti; lane 2, D. immitis; lane 3, D. repens. (A) Multiplex PCR confirmed the results of single PCR in El-Nikhila, El-Matiaa villages and Sahel Seleem district. (B) Multiplex PCR detected a positive sample of W. bancrofti (lane 1) that could not be detected by single PCR in El-Nikhila village. (C) Distribution of filarial parasites among different mosquito genera using multiplex PCR. Lane 1, Culex spp.; lane 2, Anopheles spp.; lane 3, Aedes spp. (D) Distribution of filarial parasites among different Culex species in El-Nikhila village. Lane 1, Culex pipiens; lane 2, Culex antennatus; lane 3, Culex pusillus.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4384786&req=5

f3-kjp-53-1-77: Multiplex PCR pattern on 2% ethidium bromide-stained agarose gel. Lane 1, W. bancrofti; lane 2, D. immitis; lane 3, D. repens. (A) Multiplex PCR confirmed the results of single PCR in El-Nikhila, El-Matiaa villages and Sahel Seleem district. (B) Multiplex PCR detected a positive sample of W. bancrofti (lane 1) that could not be detected by single PCR in El-Nikhila village. (C) Distribution of filarial parasites among different mosquito genera using multiplex PCR. Lane 1, Culex spp.; lane 2, Anopheles spp.; lane 3, Aedes spp. (D) Distribution of filarial parasites among different Culex species in El-Nikhila village. Lane 1, Culex pipiens; lane 2, Culex antennatus; lane 3, Culex pusillus.
Mentions: Afterwards, multiplex PCR had confirmed the single PCR results performed in the 3 localities (Fig. 3A), in fact multiplex PCR was superior to single PCR as it positively detected W. bancrofti in previously negative tested pool (Fig. 3B). The sensitivity of multiplex PCR as compared to single PCR was 100%, and the specificity was 98%. The positive predictive value was 93%, the negative predictive value was 100%, and the accuracy rate was 99% (Table 1).

Bottom Line: The technique showed 100% sensitivity and 98% specificity.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Parasitology, Faculty of Medicine, Assiut University, Assiut, Egypt.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus