@article{fdi:010058864,
  title = {{O}ptimized pan-species and speciation duplex real-time pcr assays for {P}lasmodium parasites detection in malaria vectors},
  author = {{S}andeu, {M}. {M}. and {M}oussiliou, {A}. and {M}oiroux, {N}icolas and {P}adonou, {G}. {G}. and {M}assougbodji, {A}. and {C}orbel, {V}incent and {T}uikue {N}dam, {N}icaise},
  editor = {},
  language = {{ENG}},
  abstract = {{B}ackground: {A}n accurate method for detecting malaria parasites in the mosquito's vector remains an essential component in the vector control. {T}he {E}nzyme linked immunosorbent assay specific for circumsporozoite protein ({ELISA}-{CSP}) is the gold standard method for the detection of malaria parasites in the vector even if it presents some limitations. {H}ere, we optimized multiplex real-time {PCR} assays to accurately detect minor populations in mixed infection with multiple {P}lasmodium species in the {A}frican malaria vectors {A}nopheles gambiae and {A}nopheles funestus. {M}ethods: {C}omplementary {T}aq{M}an-based real-time {PCR} assays that detect {P}lasmodium species using specific primers and probes were first evaluated on artificial mixtures of different targets inserted in plasmid constructs. {T}he assays were further validated in comparison with the {ELISA}-{CSP} on 200 field caught {A}nopheles gambiae and {A}nopheles funestus mosquitoes collected in two localities in southern {B}enin. {R}esults: {T}he validation of the duplex real-time {PCR} assays on the plasmid mixtures demonstrated robust specificity and sensitivity for detecting distinct targets. {U}sing a panel of mosquito specimen, the real-time {PCR} showed a relatively high sensitivity (88.6%) and specificity (98%), compared to {ELISA}-{CSP} as the referent standard. {T}he agreement between both methods was "excellent' (k = 0.8, {P} < 0.05). {T}he relative quantification of {P}lasmodium {DNA} between the two {A}nopheles species analyzed showed no significant difference ({P} = 0, 2). {A}ll infected mosquito samples contained {P}lasmodium falciparum {DNA} and mixed infections with {P}. malariae and/or {P}. ovale were observed in 18.6% and 13.6% of {A}n. gambiae and {A}n. funestus respectively. {P}lasmodium vivax was found in none of the mosquito samples analyzed. {C}onclusion: {T}his study presents an optimized method for detecting the four {P}lasmodium species in the {A}frican malaria vectors. {T}he study highlights substantial discordance with traditional {ELISA}-{CSP} pointing out the utility of employing an accurate molecular diagnostic tool for detecting malaria parasites in field mosquito populations.},
  keywords = {{BENIN}},
  booktitle = {},
  journal = {{P}los {O}ne},
  volume = {7},
  numero = {12},
  pages = {e52719},
  ISSN = {1932-6203},
  year = {2012},
  DOI = {10.1371/journal.pone.0052719},
  URL = {https://www.documentation.ird.fr/hor/fdi:010058864},
}