DOH95-DC-2026 ( ) 951 1 9512 31 1
500 (Beaty & Marquardt 1996) (White 2001, Estrada-Franco et al. 2003) (Russell 1998) (Kay & Farrow 2000, Johansen et al. 2003) La Crosse encephalistis ( ) the Togaviridae (genus Alphavirus)( ) Flaviridae( )Bunyaviridae (La Cross encephalitis) CDC (Moore et al. 1993) (Van der Hurk et al. 2002) (universial primers for a group of diseases in a single PCR) (spcies-specific primers) (Kuno 1998, Bronzoni et al. 2005) RNA Real time PCR Anderson et al. 2004, Falco et al. 2002, Mboera et al. 2000 Russell & Hunter 2005 CDC ( ) ( / ) (Modified from Beaty & 2
Marquart, 1996Moore et al., 1993CDC, 2003) Flaviviridae 3-12% 0% 30-40% 7%( 4-20% Aedes aegypti Aedes albopictus Culex annulirostris Culex gelidus Culex vishinui Culex fuscocephalus Culex tritaeniorhyncuhs ) Aedes spp. (4) Aedes albopictus Aedes triseriatus Aedes vexans Anopheles spp.(6) Coquillettidia sp.(1) Culex spp. (8) Culex pipiens Culex quinquefasciatus Culex restuans Culex salinarius Culex tarsalis Culex nigripalpus Dinocerites sp.(1) Ochlerotatus spp.(15) Culiseta spp. (2) Orthopodomyia sp.(1) Psorophora spp. (4) Uranotaenia sp.(1) Culex restuans Culex salinarius Culex nigripalpus Culex pipiens complex Culex tarsalis 20-70% Culex annulirostris 5-20% Aedes aegypti Aedes africanus Aedes simpsoni Haemagogus spp. Togaviridae 50-75% 5-10% 0.1-20% 0% Aedes albopictus Aedes canadensis Aedes sollictians Aedes vexans Coquillettidia perturbans Culex nigripalpus Culex salinarius Culisera melanura Aedes melanimon Culex tarsalis Aedes taeniorhychus Culex spp.(30). Culex annulirostris Ochlerotatus(Ae.) vigilax Aedes procax Aedes funereus Aedes aegypti Aedes albopictus Culex vishinui Culex fuscocephalus Culex tritaeniorhyncuhs Aedes albopictus Aedes aegypti Aedes vexans Culex quinqefasciatus Ochlerotatus dorsalis Ochlerotatus japonicus Culex quinqefasciatus Aedes aegypti Aedes albopictus Aedes vexans Culex quinquefasciatus Ochlerotatus vigilax 3
Bunyaviridae La Crosse encephalistis Aedes aegypti Aedes aegypti 1% Small mammals ( Tamias striatus) 0% Aedes Canadensis Aedes communis Aedes dorsalis Aedes melanimon Aedes stimulans Aedes triseriatus Culiseta inornata Floodwater Aedes spp Culex pipiens. Culex quinquefasciatus? () () RNA 4
() (mosquito-borne arboviral encephalitis) duplex reverse transcription PCR RNA cdna Alphavirus flavivirus forward cm3w ( 5 -YAGAGCDTTTTCGCAYSTRGCHW-3 )reverse cm3w (5 -ACATRAANKGNGTNGTCRAANCCDAYCC-3 ) Alphavirus 434bp forward FG1(5 -TCAAGGAACTCCACACATGAGATGTACT-3 )reverse FG25 -GTGTCCCATCCTGCTGTGTCATCAGCATACA-3 Flavivirus NS5 958bp 50 C 30 95 C 15 35 94 C 1 denature 53 C 1 (annealing)72 C 2 (elongation) 72 C 5 (extension)2agarose gel QIAquick gel extraction kit (QIAGEN) multiplex nested PCR, M-N-PCR Alphavirus Flavivirus 94 C 1 denature 53 C 1 (annealing)72 C 2 (elongation) 25 72 C 5 (extension) 2agarose gel 5
(A) (B) (C) 6
7
8
B 9
18,425 14,424 56 11343 724 46159 164132 266 206 10842 8 2 231 4 862 ( 95 6 22 UD 50 95 9 15 UD 50 Alpha 866 95 351 14424 56 14480 2 57 1134 3 1137 0 30 724 102 826 0 94 461 59 520 0 78 164 132 296 0 20 266 1 267 0 12 206 27 233 0 41 108 42 150 0 18 143 8 151 0 10 60 2 62 0 8 57 1 58 0 16 33 0 33 0 8 29 0 29 0 12 18 5 23 0 5 15 6 21 0 13 8 9 17 0 12 15 0 15 0 8 13 1 14 0 11 13 1 14 0 5 11 2 13 0 7 11 4 15 0 2 12 1 13 0 9 10 2 12 0 6 12 0 12 0 4 11 0 11 0 8 8 2 10 0 4 4 1 5 0 2 3 1 4 0 2 0 2 2 0 10
2 1 1 2 0 2 1 1 2 0 1 2 0 2 0 1 1 0 1 0 1 1 0 1 0 1 2 0 2 0 1 1 0 1 0 1 1 0 1 0 1 0 1 1 0 1 1 0 1 0 866 17,970 473 18,425 2 833 Ct 40 Tm 80 4 PCR 3 958 bp 2 alpha 866 Ct 40 Tm 80 11 434 bp dimers Real time RT-PCR alpha Real time RT-PCR 11
866 PCR 4 11 1 11 2 MIR 0.11% pool UD CDC 2 50 CDC CDC UD UD 2004 Beaty BJ, and Marquardt WC. 1996. The biology of disease vectors. The University Press of Colorado, Colorado, USA, 85-97pp. Boom AK, Lindsay MD, Wright AE, Smith DW, Mackenzie JS. 2003. Epizootic activity of Murray Valley encephalitis and Kunjin viruses in an aboriginal community in the southeast Kimberley region of Western Australia: results of mosquito fauna and virus isolation studies. Amer. J. Trop. Med. & Hygiene 69: 277-283. Center for Disease Control. Epidemic/Epizootic West Nile Virus in the United States: Guidelines for surveillance, prevention, and control. 2003, 75pp. Estrada-Franco JG. Navarro-Lopez R. Beasley DWC. Coffey L. Carrara AS. Da Rosa AT. Clements T. Wang E. Ludwig GV. Cortes AC. Ramirez PP. Tesh RB. Barrett ADT. Weaver SC. 2003. West nile virus in Mexico: Evidence of widespread circulation since July 2002. Emerging Infectious Diseases 9:1604-1607. Johansen CA, Nisbet DJ, Zborowski P, van den Hurk AF, Ritchie SA, Mackenzie JS. Flavivirus isolations from mosquitoes collected from western Cape York Peninsula, Australia, 1999-2000. J. Amer. Mosq. Control Assoc. 19:392-396. Kay BH, and Farrow RA. 2000. Mosquito (Diptera:Culicidae) dispersal: implications fro the epidemiology of Japanese and Murray Valley Encephalitis Viruses in Australia. J. Med. Entomol. 37: 797-801. 12
Mcnelly JR. 1989. The CDC trap as a special monitoring tool. Moore CG. Mclean RG. Mitchell CJ. Nasci RS. Tsai TF. Calisher CH. Marfin AA. Moore PS. Gubler DJ. 1993. Guidelines for arbovirus surveillance programs in the United States. CDC. 81 pp. Russell, RC. 1998. Vectors vs. human in Australia-who is on top down under? An update on vector-borne disease and research on vectors in Australia. J. Vector Eco. 23:1-46. Spencer JD, Azoulas J, Broom AK, Buick TD, Currie B, Daniels PW, Doggett SL, Hapgood GD, Jarrett PJ, Lindsay MD, Lloyd G, Mackenzie JS, Merianos A, Moran RJ, Ritchie SA, Russell RC, Smith DW, Stenhouse FO, Whelan PI. 2001. Murray valley encephalitis virus surveillance and control initiatives in Australia. National Arbovirus Advisory Committee of the Communicable Diseases Network Australia. Communicable Diseases Intelligence 25:33-47. White DJ, Kramer LD, Backenson PB, Lukacik G, Johnson G, Oliver J, Howard JJ, Means RG, Eidson M, Gotham I, Kulasekera V, Campbell S. Mosquito surveillance and polymerase chain reaction detection of west nile virus, New York State. Emerging Infectious diseases 7:643-649, 2001. 13