106                                                                                                             S.BONNETET...
lXA.SMODIUM     DETECTION       IN MOSQUITOES                                                                             ...
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2000 infectivity of malaria vector mosquitoes

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2000 infectivity of malaria vector mosquitoes

  1. 1. 106 S.BONNETETAL falciparum. Transactions of theRoyal Socieg of TropicalMedicine Roeffen, W., Geeraedts, F., Eling, W., Beckers, I’., Wizel, B., and Hygiene, 74,738-742. Kumar, N., Lensen, T. & Sauerwein, R. (1995). Transmis-Graves, I’. M., Burkot, T. R., Carter, R., Cattani, J. A., Lagog, sion blockade of Plasmodium falciparum malaria by anti- M., Parker, J., Brabin, B. J., Gibson, F. D., Bradley, D. J. & Pfs230-specific antibodies is isotype dependent. Infection AlDers, M. P. (1988). Measurement of malaria infectivitv of and Immunity, 63,467-471. himan populations to mosquitoes in the Madang area, Papua Rutledge, L. C., Ward, R. A. & Gould, D. J. (1964). Studies on New Guinea. Parasitology, 96,251-263. the feeding response of mosquitoes to nutritive solutions in aJeffery, G. & Eyles, D. E. (1955). Infectivity to mosquitoes of new membrane feeder. Mosquito New!, 24, 407-419. Plasmodium falcioarum as related to aametocvte densitv and Tchuinkam, T., Mulder, B., Dechermg, K., Stoffels, H., duration of ‘mfection. American Jot&al of Tiopical Me&&e Verhave, J. P., Cot, M., Carnevale, I’., Meuwissen, J. H. E. and Hygiene, 4, 781-789. T. & Robert, V. (1993). Experimental infections of AnophelesKaslow, D. C. (1993). Transmission-blocking immunity gambiae with Plasmodium falciparum of naturally infected against malaria and other vector-borne diseases. Current gametocvte carriers in Cameroon: factors influencing the Opinion in Immunology, 5, 557-565. hfectiv& to mosquitoes. Tropical Medicine and Parasitology,Lensen, A., Vandruten, J., Bolmer, M., Vangemert, G., Eling, 44,271-276. W. & Sauerwein, R. (1996). Measurement by membrane Vanderberg, J. P. & Gwadz, R. W. (1980). The transmission by feeding of reduction in Plasmodium falciparum transmission mosquitoes of plasmodia in the laboratory. In: Malaria, induced by endemic sera. Transactions of the Royal Society of Kreier, J. I’. (editor). New York: Academic Press, pp. 154- Tropical Medicine and Hygiene, 90, 20-22. 218.Muirhead-Thomson, R. C. (1957). The malarial infectivity of Yoeli, M. (1938). Note on the experimental infection of an African village population to mosquitoes (Anopheles gam- Anopheles elutus with Plasmodium falciparum by feeding biae) American Journal of Tropical Medicine and Hygiene, 6, through a prepared animal membrane. Rivista diMaZariologia, 971-979. 17,62-66.Ponnudurai, T., Lensen, A. H. W., Gemert Van, G. J. A., Bensink, M. P. E., Bolmer, M. & Meuwissen, J. H. E. T. (1989). Infectivity of cultured Plasmodium falciparum game- Received 25 May 1999; revised 18 3;11y 1999; accepted for tocytes to mosquitoes. Parasitology,98, 165-173. publication 3 August 1999TRANSACTIONS OF THE ROYAL SOCIETY OF TROPICAL MEDICINE AND HYGIENE (2000) 94,106-107 Amapi, Rondbnia and Roraima States). The same num-1 Short Report 1 ber of mosquitoes known to be negative for humanI I malaria parasites was also tested. The source of PZusmodium DNA was the same materialInfectivity of malaria vector used for the ELISA test [triturated mosquitoes-head and thorax ground in a blocking buffer containing 0.05%mosquitoes: correlation of positivity nonidet P-40 (WIRTZ et al., 1987)], either 20 @spottedbetween ELBA and PCR-ELISA tests on a glass fibre membrane (GFM) prepared for PCR, using l/8 of the spot as DNA source directly into the PCR mixture as described by WARHURST et al. (1991), orMarinete M. l%voa’*, Ricardo L. D. Machado’, extracted as follows: 40 & of ELISA solution wasMaria N. 0. Segura’, Giselle M. R. Vianna’, centrifuged for 10 min at 22 500 g, the pellet was lysedAdenildo S. Vasc&celds’ and Jan E. Conn3 ‘Pro: by adding 25 &of lysis buffer and incubating at 65°C forwama de Malhia. Instituto Evandro ChaPaslFAWMS. 30 min, afterwards adding potassium acetate and placingk. Almirante Bakroso, 492, 66.090-000, -Bel&m, Park, on ice for at least 60 min. The isolated DNA (obtained byBrazil; ‘Institute Oswald0 Cruz, FIOCRUZ, Av. Brasil, ethanol precipitation) was dissolved in 15 pL of TE-436.5, 210 45-900, Rio deyaneiro, RJ, Brazil; 3Department buffer containing ribonuclease WILSON et al., 1998).of Biology, Marsh Life Sciences Building, University of The DNA was amplified as described by MACHADO et al.Vermont, Burlington, VT, 054050460, USA (1998) using primer sequence, concentrations and reac- tion conditions indicated by OLIVEIRA et al. (1995). For the identification of the human malaria parasites we usedKeywords: malaria transmission, anopheline mosquitoes, in- the liquid-phase, non-isotopic hybridization ELISAfectivity, ELISA, PCR-ELISA, Brazil technique, following the protocol of OLIVEIRA et al. (1995). For negative controls we used distilled water, Studies on the infectivity of malaria vector mosquitoes male anopheline and culicine mosquitoes, and humanusing the enzyme-linked immunosorbent assay (ELISA) DNA. Positive controls included strain Kl of I? fulcipar-described by WIRTZ et al. (1987) have been carried out urn, and mosquitoes experimentally infected with l?worldwide for several years. SOMBOON et al. (1993) falciparum and l? vivax.reported false-positive results for the ELISA associated Our PCR results confirmed the ELISA test results forwith bovine and swine blood. In order to avoid these all positive and all negative mosquitoes, and in 5 (15.6%)false-positive results it is advisable either to use only the of the positive mosquitoes (3 An. albitursis and 2 An.anterior part of the mosquito (head and thorax) which, darlingi) the PCR-ELISA technique detected othernormally, is not contaminated by the ingested animal species of human Plasmodium that were not found byblood (WIRTZ et al., 1987). or to confirm the ELISA the ELISA test alone (Table). The DNA source wasresult by another method’such as polymerase chain obtained only by DNA extraction, which means that thereaction (PCR). GFM technique is not applicable for this tvpe of material. We have carried out the PCR-ELISA to confirm the These results indicate-&at the PCR-&ISA is moredetection of human malaria parasites in mosquitoes sensitive than a simnle ELISA test which. however. stillalready recorded as positive by ELISA alone. Thirty remains a very goodand useful tool for testing mo&itotwo such mosquitoes were tested, belonging to different infectivity.species of the genus Anopheles and collected during fieldtrips to different areas of the Amazonia Region (Pari, Acknowledgements We thank the staff of the Malaria Entomology Laboratory at the Evandro Chagas Institute for technical assistance and*Author for correspondence; fax +55 91226 1284 or 2114417. Professor Ralph Lainson for reviewing the manuscript. This
  2. 2. lXA.SMODIUM DETECTION IN MOSQUITOES 107 Table. Comparison between ELISA and PCR-ELISA results for human malaria parasites in Anopheles mosquitoes from several areas ofthe Amazon region in Brazil Species State ELISA PCR-ELISA An. (NY.) albitarsis Roraima PVIPf PvllwPm An. (Nys.) albitarsis PvlPj An. (Nys.) albitarsis Roraima filpf An. (N&.j albitarsis Roraima PvlPf An. fNvs. ) braziliensis Roraima PvlPf An. &$.j nuneztovari Roraima Pvlps An. (Nys.) darlingi Rondbnia An. (Nys.) darlingi Rondhnia g An. (Nys.) albitarsis Amap y&k An. (Nys.) albitarsis Amapi An. (Nys.) albitarsis Amapi rym An. (Nys.) albitarsis Amaph Pv An. (Nys.) albitarsis AmapL l% An. (Nys.) albitarsis Amapk Pm An. (Nys.) albitarsis Amapi Pv An. (Nys.) braziliensis Amapi Pv An. (Nys.) darlingi Amapi An. (Nys.) darlingi Amapi x An. (Nys.) aquasalis ParP An. (Nys.) aquasalis Pari An. (Nys.) aquasalis Pari An. (Nys.) darlingi Pari Pv An. (Nys.) darling’ Pari PV An. (Nys.) darlingi Pari An. (Nys.) darlingi Pari An. (Nys.) nuneztovari Pari An. (Nys.) nuneztovari Pari An. (Nys.) nuneztovari Pari An. (Nys.) nuneztovan’ Pari An. (Nys.) nuneztovari Pa& An. (Nys.) nuneztovari Pari Pm An. (Nys.) nuneztovari Pari Pm Pv, Plasmodium vivax; pf, l? falciparum; Pm, I? malariae.study received financial support from a National Institute of Warhurst, D. C., Awad-el-Karien, F. M. & Miles, M. A. (199 1).Health Grant (A140116) to J.E.C. and Evandro Chagas In- Simplified preparation of malaria blood samples for polymer-stitute/FNS. ase chain reaction. Lancet, 337, 303-304. Wilson, M. D., Ofosu-Okyere, A., Okoli, A. U., McCall, P. J. &References Snounou, G. (1998). Direct comparison of microscopy andMachado, R. L. D., Garret, D. O., Adagu, I. S., Warhurst, D. C. polymerase chain reaction for the detection of Plasmodium & I%voa, M. M. (1998). Simolified diagnosis of malaria sporozoites in salivary glands of mosquitoes. Transactions of infection: GFM/PC&E&A, a-simplified-nucleic acid am- the Royal Society of Tropical Medicine and Hvpiene, 92, plification technique by PClUELISA. Revista do Institute de 482-483. Medicina Tropical de SLio Paulo, 40, 333-334. Wirtz, R. A., Zavala, F., Charoenvit, Y., Campell, G. H.,Oliveira, D. A., Holloway, B. P., Durigon, E. L., Collins, W. E. Burkot, T. R., Scheneider, I., Esser, K. M., Beaudoin, R. L. & Lal, A. A. (1995). Polymerase chain reaction and liquid- & Andr& R. G. (1987). Comparative testing of monoclonal phase, nonisotopic hybridization for species-specific and antibody against Plasmodiumfalciparum sporozoite for ELISA sensitive detection of malaria infection. American Journal of development. Bulletin of the World Health Organization, 65, Tropical Medicine and Hygiene, 52, 139- 144. 39-45.Somboon, I’., Morakote, N., Koottathep, S. & Trisanarom, U. (1993). Detection of sporozoites of Plasmodium vivax and Plasmodium falciparum in mosquitoes by ELISA: false posi- tivity associated with bovine and swine blood. Transactions of the Royal Society of Tropical Medicine and Hygiene, 87, Received ISJune 1999; revised 28 September 1999; accepted 322-324. for publication 1 October 1999

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