Author:
- INTRODUCTION
- MATERIALS AND METHODS
- Mosquito material
- Nucleic acid isolation
- ITS1 PCR amplification and sequencing
- Real-time PCR primer and probe design
- Real-time PCR amplifications
- Synthetic primer and probe controls
- Validation of real-time PCR assays
- Assessment of the real-time PCR assays using field collected samples
- Evaluation of FTA cards and filter paper for sample preservation
- RESULTS
- PCR amplification and sequencing of ITS1
- Synthetic control development
- Sensitivity of real-time PCR assays
- Specificity of real-time PCR assays
- Identification of different life stages and mixed pools
- Assessment of real-time PCR assays using field collected samples
- Evaluation of FTA cards for sample preservation
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Benedict MQ, Levine RS, Hawley WA, Lounibos LP, 2007. Spread of the tiger: global risk of invasion by the mosquito Aedes albopictus. Vector Borne Zoonotic Dis 7 :76ā85.
-
2
Gratz NG, 2004. Critical review of the vector status of Aedes albopictus. Med Vet Entomol 18 :215ā227.
-
3
Almeida AP, Baptista SS, Sousa CA, Novo MT, Ramos HC, Panella NA, Godsey M, SimoĢes MJ, Anselmo ML, Komar N, Mitchell CJ, Ribeiro H, 2005. Bioecology and vectorial capacity of Aedes albopictus (Diptera: Culicidae) in Macao, China, in relation to dengue virus transmission. J Med Entomol 42 :419ā428.
-
4
Effler PV, Pang L, Kitsutani P, Vorndam V, Nakata M, Ayers T, Elm J, Tom T, Reiter P, Rigau-Perez JG, Hayes JM, Mills K, Napier M, Clark GG, Gubler DJ, 2005. Dengue fever, Hawaii, 2001ā2002. Emerg Infect Dis 11 :742ā749.
-
5
Reiter P, Fontenille D, Paupy C, 2006. Aedes albopictus as an epidemic vector of chikungunya virus: another emerging problem? Lancet Infect Dis 6 :463ā464.
-
6
Vazeille M, Moutailler S, Coudrier D, Rousseaux C, Khun H, Huerre M, Thiria J, Dehecq JS, Fontenille D, Schuffenecker I, Despres P, Failloux AB, 2007. Two Chikungunya isolates from the outbreak of La Reunion (Indian Ocean) exhibit different patterns of infection in the mosquito, Aedes albopictus. PLoS ONE 2 :e1168.
-
7
Rezza G, Nicoletti L, Angelini R, Romi R, Finarelli AC, Panning M, Cordioli P, Fortuna C, Boros S, Magurano F, Silvi G, Angelini P, Dottori M, Ciufolini MG, Majori GC, Cassone A, 2007. Infection with chikungunya virus in Italy: an outbreak in a temperate region. Lancet 370 :1840ā1846.
-
8
Whelan P, Hapgood G, 2001. A mosquito survey of Dili, East Timor, and implications for disease control. Arbovirus Res Aust 8 :405ā416.
-
9
Schoenig E, 1972. Distribution of 3 species of Aedes (Stegomyia) carriers of virus diseases on the main island of Papua and New Guinea. Philipp Sci 9 :61ā82.
-
10
Kay BH, Prakash G, Andre RG, 1995. Aedes albopictus and Aedes (Stegomyia) species in Fiji. J Am Mosq Control Assoc 11 :230ā234.
-
11
Elliot SA, 1980. Aedes albopictus in the Solomon and Santa Cruz islands, South Pacific. Trans R Soc Trop Med Hyg 74 :747ā748.
-
12
Cooper RD, Waterson DGE, Kupo M, Sweeney AW, 1994. Aedes albopictus (Skuse) (Diptera: Culicidae) in the Western Province of Papua New Guinea and the threat of its introduction to Australia. J Aust Entomol Assoc 33 :115ā116.
-
13
Russell RC, Williams CR, Sutherst RW, Ritchie SA, 2005. Aedes (Stegomyia) albopictusāa dengue threat for southern Australia. Comm Dis Intell 29 :296ā298.
-
14
Mitchell CJ, Gubler DJ, 1987. Vector competence of geographic strains of Aedes albopictus and Aedes polynesiensis and certain other Aedes (Stegomyia) mosquitoes for Ross River virus. J Am Mosq Control Assoc 3 :142ā147.
-
15
Ritchie SA, Moore P, Carruthers M, Williams C, Montgomery B, Foley P, Ahboo S, van den Hurk AF, Lindsay MD, Cooper B, Beebe N, Russell RC, 2006. Discovery of a widespread infestation of Aedes albopictus in the Torres Strait, Australia. J Am Mosq Control Assoc 22 :358ā365.
-
16
Rai KS, Pashley DP, Munstermann LE, 1982. Genetics of speciation in Aedine mosquitoes. Steiner WM, Tabachnick WJ, Rai KS, Narang S, eds. Recent Developments in the Genetics of Insect Disease Vectors. Champaign, Illinois: Stipes Publishers, 84ā129.
-
17
Lee DJ, Hicks MM, Griffiths M, Debenham ML, Bryan JH, Russell RC, Geary M, Marks EN, 1987. The Culicidae of the Australasian Region, Vol. 4. Entomology Monograph No. 2. Canberra: Australian Government Publishing Service Press.
-
18
Huang YM, 1972. The subgenus Stegomyia of Aedes in Southeast Asia. IāThe Scutellaris group of species. Contrib Am Entomol Inst 9 :1ā109.
-
19
Lamche GD, Whelan PI, 2003. Variability of larval identification characters of exotic Aedes albopictus (Skuse) intercepted in Darwin, Northern Territory. Comm Dis Intell 27 :105ā109.
-
20
Beebe NW, Whelan PI, van den Hurk AF, Ritchie SA, Corcoran S, Cooper RD, 2007. A polymerase chain reaction-based diagnostic to identify larvae and eggs of container mosquito species from the Australian region. J Med Entomol 44 :376ā380.
-
21
Sinclair D, 1992. The distribution of Aedes aegypti and dengue in Queensland, 1990āJune 30, 1992. Arbovirus Res Aust 6 :323.
-
22
Beebe NW, Whelan PI, van den Hurk A, Ritchie SA, Cooper RD, 2005. Genetic diversity of the dengue vector Aedes aegypti in Australia and implications for future surveillance and mainland incursion monitoring. Comm Dis Intell 29 :299ā304.
-
23
Beebe NW, Cooper RD, Foley DH, Ellis JT, 2000. Populations of the south-west Pacific malaria vector Anopheles farauti s.s. revealed by ribosomal DNA transcribed spacer polymorphisms. Heredity 84 :244ā253.
-
24
Heid CA, Stevens J, Livak KJ, Williams PM, 1996. Real time quantitative PCR. Genome Res 6 :986ā994.
-
25
Livak KJ, Flood SJA, Marmaro J, Giusti W, Deetz K, 1995. Oligonucleotides with fluorescent dyes at opposite ends provide a quenched probe system useful for detecting PCR product and nucleic acid hybridization. PCR Methods Appl 4 :357ā362.
-
26
Smith G, Smith I, Harrower B, Warrilow D, Bletchly C, 2006. A simple method for preparing synthetic controls for conventional and real-time PCR for the identification of endemic and exotic disease agents. J Virol Methods 135 :229ā234.
-
27
Rutledge RG, CoĢteĢ C, 2003. Mathematics of quantitative kinetic PCR and the application of standard curves. Nucleic Acids Res 31 :e93.
-
28
Bass C, Williamson MS, Wilding CS, Donnelly MJ, Field LM, 2007. Identification of the main malaria vectors in the Anopheles gambiae species complex using a TaqMan real-time PCR assay. Malar J 6 :155.
-
29
Marks EN, 1980. Mosquitoes (Diptera: Culicidae) of Cape York Peninsula, Australia. Stevens NC, Bailey A, eds. Contemporary Cape York Peninsula. Brisbane: The Royal Society of Queensland, 59ā76.
-
30
Sanogo YO, Kim CH, Lampman R, Novak RJ, 2007. A real-time TaqMan polymerase chain reaction for the identification of Culex vectors of West Nile and Saint Louis encephalitis viruses in North America. Am J Trop Med Hyg 77 :58ā66.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 41 | 41 | 6 |
| Full Text Views | 424 | 139 | 0 |
| PDF Downloads | 247 | 71 | 0 |
Rapid Identification of Aedes albopictus, Aedes scutellaris, and Aedes aegypti Life Stages Using Real-time Polymerase Chain Reaction Assays
Lydia A. Hill
Lydia A. Hill
School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland, Australia; Tropical Population Health Unit Network, Queensland Health, Cairns, Queensland, Australia; Medical Entomology, Centre for Disease Control, Department of Health and Community Services, Darwin, Northern Territory, Australia; Australian Army Malaria Institute, Gallipoli Barracks, Enoggera, Queensland, Australia; Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, Queensland, Australia
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Joseph B. Davis
Joseph B. Davis
School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland, Australia; Tropical Population Health Unit Network, Queensland Health, Cairns, Queensland, Australia; Medical Entomology, Centre for Disease Control, Department of Health and Community Services, Darwin, Northern Territory, Australia; Australian Army Malaria Institute, Gallipoli Barracks, Enoggera, Queensland, Australia; Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, Queensland, Australia
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George Hapgood
George Hapgood
School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland, Australia; Tropical Population Health Unit Network, Queensland Health, Cairns, Queensland, Australia; Medical Entomology, Centre for Disease Control, Department of Health and Community Services, Darwin, Northern Territory, Australia; Australian Army Malaria Institute, Gallipoli Barracks, Enoggera, Queensland, Australia; Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, Queensland, Australia
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Peter I. Whelan
Peter I. Whelan
School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland, Australia; Tropical Population Health Unit Network, Queensland Health, Cairns, Queensland, Australia; Medical Entomology, Centre for Disease Control, Department of Health and Community Services, Darwin, Northern Territory, Australia; Australian Army Malaria Institute, Gallipoli Barracks, Enoggera, Queensland, Australia; Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, Queensland, Australia
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Greg A. Smith
Greg A. Smith
School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland, Australia; Tropical Population Health Unit Network, Queensland Health, Cairns, Queensland, Australia; Medical Entomology, Centre for Disease Control, Department of Health and Community Services, Darwin, Northern Territory, Australia; Australian Army Malaria Institute, Gallipoli Barracks, Enoggera, Queensland, Australia; Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, Queensland, Australia
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Scott A. Ritchie
Scott A. Ritchie
School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland, Australia; Tropical Population Health Unit Network, Queensland Health, Cairns, Queensland, Australia; Medical Entomology, Centre for Disease Control, Department of Health and Community Services, Darwin, Northern Territory, Australia; Australian Army Malaria Institute, Gallipoli Barracks, Enoggera, Queensland, Australia; Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, Queensland, Australia
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R. D. Cooper
R. D. Cooper
School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland, Australia; Tropical Population Health Unit Network, Queensland Health, Cairns, Queensland, Australia; Medical Entomology, Centre for Disease Control, Department of Health and Community Services, Darwin, Northern Territory, Australia; Australian Army Malaria Institute, Gallipoli Barracks, Enoggera, Queensland, Australia; Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, Queensland, Australia
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Andrew F. van den Hurk
Andrew F. van den Hurk
School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland, Australia; Tropical Population Health Unit Network, Queensland Health, Cairns, Queensland, Australia; Medical Entomology, Centre for Disease Control, Department of Health and Community Services, Darwin, Northern Territory, Australia; Australian Army Malaria Institute, Gallipoli Barracks, Enoggera, Queensland, Australia; Virology, Forensic and Scientific Services, Queensland Health, Coopers Plains, Queensland, Australia
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In 2005, a widespread infestation of Aedes albopictus was discovered in the Torres Strait, the region between northern Australia and New Guinea. To contain this species, an eradication program was implemented in 2006. However, the progress of this program is impeded by the difficulty of morphologically separating Ae. albopictus larvae from the endemic species Aedes scutellaris. In this study, three real-time TaqMan polymerase chain reaction assays that target the ribosomal internal transcribed spacer 1 region were developed to rapidly identify Aedes aegypti, Ae. albopictus, and Ae. scutellaris from northern Australia. Individual eggs, larvae, pupae, and adults, as well as the species composition of mixed pools were accurately identified. The assay method was validated using 703 field-collected specimens from the Torres Strait.
- INTRODUCTION
- MATERIALS AND METHODS
- Mosquito material
- Nucleic acid isolation
- ITS1 PCR amplification and sequencing
- Real-time PCR primer and probe design
- Real-time PCR amplifications
- Synthetic primer and probe controls
- Validation of real-time PCR assays
- Assessment of the real-time PCR assays using field collected samples
- Evaluation of FTA cards and filter paper for sample preservation
- RESULTS
- PCR amplification and sequencing of ITS1
- Synthetic control development
- Sensitivity of real-time PCR assays
- Specificity of real-time PCR assays
- Identification of different life stages and mixed pools
- Assessment of real-time PCR assays using field collected samples
- Evaluation of FTA cards for sample preservation
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Benedict MQ, Levine RS, Hawley WA, Lounibos LP, 2007. Spread of the tiger: global risk of invasion by the mosquito Aedes albopictus. Vector Borne Zoonotic Dis 7 :76ā85.
-
2
Gratz NG, 2004. Critical review of the vector status of Aedes albopictus. Med Vet Entomol 18 :215ā227.
-
3
Almeida AP, Baptista SS, Sousa CA, Novo MT, Ramos HC, Panella NA, Godsey M, SimoĢes MJ, Anselmo ML, Komar N, Mitchell CJ, Ribeiro H, 2005. Bioecology and vectorial capacity of Aedes albopictus (Diptera: Culicidae) in Macao, China, in relation to dengue virus transmission. J Med Entomol 42 :419ā428.
-
4
Effler PV, Pang L, Kitsutani P, Vorndam V, Nakata M, Ayers T, Elm J, Tom T, Reiter P, Rigau-Perez JG, Hayes JM, Mills K, Napier M, Clark GG, Gubler DJ, 2005. Dengue fever, Hawaii, 2001ā2002. Emerg Infect Dis 11 :742ā749.
-
5
Reiter P, Fontenille D, Paupy C, 2006. Aedes albopictus as an epidemic vector of chikungunya virus: another emerging problem? Lancet Infect Dis 6 :463ā464.
-
6
Vazeille M, Moutailler S, Coudrier D, Rousseaux C, Khun H, Huerre M, Thiria J, Dehecq JS, Fontenille D, Schuffenecker I, Despres P, Failloux AB, 2007. Two Chikungunya isolates from the outbreak of La Reunion (Indian Ocean) exhibit different patterns of infection in the mosquito, Aedes albopictus. PLoS ONE 2 :e1168.
-
7
Rezza G, Nicoletti L, Angelini R, Romi R, Finarelli AC, Panning M, Cordioli P, Fortuna C, Boros S, Magurano F, Silvi G, Angelini P, Dottori M, Ciufolini MG, Majori GC, Cassone A, 2007. Infection with chikungunya virus in Italy: an outbreak in a temperate region. Lancet 370 :1840ā1846.
-
8
Whelan P, Hapgood G, 2001. A mosquito survey of Dili, East Timor, and implications for disease control. Arbovirus Res Aust 8 :405ā416.
-
9
Schoenig E, 1972. Distribution of 3 species of Aedes (Stegomyia) carriers of virus diseases on the main island of Papua and New Guinea. Philipp Sci 9 :61ā82.
-
10
Kay BH, Prakash G, Andre RG, 1995. Aedes albopictus and Aedes (Stegomyia) species in Fiji. J Am Mosq Control Assoc 11 :230ā234.
-
11
Elliot SA, 1980. Aedes albopictus in the Solomon and Santa Cruz islands, South Pacific. Trans R Soc Trop Med Hyg 74 :747ā748.
-
12
Cooper RD, Waterson DGE, Kupo M, Sweeney AW, 1994. Aedes albopictus (Skuse) (Diptera: Culicidae) in the Western Province of Papua New Guinea and the threat of its introduction to Australia. J Aust Entomol Assoc 33 :115ā116.
-
13
Russell RC, Williams CR, Sutherst RW, Ritchie SA, 2005. Aedes (Stegomyia) albopictusāa dengue threat for southern Australia. Comm Dis Intell 29 :296ā298.
-
14
Mitchell CJ, Gubler DJ, 1987. Vector competence of geographic strains of Aedes albopictus and Aedes polynesiensis and certain other Aedes (Stegomyia) mosquitoes for Ross River virus. J Am Mosq Control Assoc 3 :142ā147.
-
15
Ritchie SA, Moore P, Carruthers M, Williams C, Montgomery B, Foley P, Ahboo S, van den Hurk AF, Lindsay MD, Cooper B, Beebe N, Russell RC, 2006. Discovery of a widespread infestation of Aedes albopictus in the Torres Strait, Australia. J Am Mosq Control Assoc 22 :358ā365.
-
16
Rai KS, Pashley DP, Munstermann LE, 1982. Genetics of speciation in Aedine mosquitoes. Steiner WM, Tabachnick WJ, Rai KS, Narang S, eds. Recent Developments in the Genetics of Insect Disease Vectors. Champaign, Illinois: Stipes Publishers, 84ā129.
-
17
Lee DJ, Hicks MM, Griffiths M, Debenham ML, Bryan JH, Russell RC, Geary M, Marks EN, 1987. The Culicidae of the Australasian Region, Vol. 4. Entomology Monograph No. 2. Canberra: Australian Government Publishing Service Press.
-
18
Huang YM, 1972. The subgenus Stegomyia of Aedes in Southeast Asia. IāThe Scutellaris group of species. Contrib Am Entomol Inst 9 :1ā109.
-
19
Lamche GD, Whelan PI, 2003. Variability of larval identification characters of exotic Aedes albopictus (Skuse) intercepted in Darwin, Northern Territory. Comm Dis Intell 27 :105ā109.
-
20
Beebe NW, Whelan PI, van den Hurk AF, Ritchie SA, Corcoran S, Cooper RD, 2007. A polymerase chain reaction-based diagnostic to identify larvae and eggs of container mosquito species from the Australian region. J Med Entomol 44 :376ā380.
-
21
Sinclair D, 1992. The distribution of Aedes aegypti and dengue in Queensland, 1990āJune 30, 1992. Arbovirus Res Aust 6 :323.
-
22
Beebe NW, Whelan PI, van den Hurk A, Ritchie SA, Cooper RD, 2005. Genetic diversity of the dengue vector Aedes aegypti in Australia and implications for future surveillance and mainland incursion monitoring. Comm Dis Intell 29 :299ā304.
-
23
Beebe NW, Cooper RD, Foley DH, Ellis JT, 2000. Populations of the south-west Pacific malaria vector Anopheles farauti s.s. revealed by ribosomal DNA transcribed spacer polymorphisms. Heredity 84 :244ā253.
-
24
Heid CA, Stevens J, Livak KJ, Williams PM, 1996. Real time quantitative PCR. Genome Res 6 :986ā994.
-
25
Livak KJ, Flood SJA, Marmaro J, Giusti W, Deetz K, 1995. Oligonucleotides with fluorescent dyes at opposite ends provide a quenched probe system useful for detecting PCR product and nucleic acid hybridization. PCR Methods Appl 4 :357ā362.
-
26
Smith G, Smith I, Harrower B, Warrilow D, Bletchly C, 2006. A simple method for preparing synthetic controls for conventional and real-time PCR for the identification of endemic and exotic disease agents. J Virol Methods 135 :229ā234.
-
27
Rutledge RG, CoĢteĢ C, 2003. Mathematics of quantitative kinetic PCR and the application of standard curves. Nucleic Acids Res 31 :e93.
-
28
Bass C, Williamson MS, Wilding CS, Donnelly MJ, Field LM, 2007. Identification of the main malaria vectors in the Anopheles gambiae species complex using a TaqMan real-time PCR assay. Malar J 6 :155.
-
29
Marks EN, 1980. Mosquitoes (Diptera: Culicidae) of Cape York Peninsula, Australia. Stevens NC, Bailey A, eds. Contemporary Cape York Peninsula. Brisbane: The Royal Society of Queensland, 59ā76.
-
30
Sanogo YO, Kim CH, Lampman R, Novak RJ, 2007. A real-time TaqMan polymerase chain reaction for the identification of Culex vectors of West Nile and Saint Louis encephalitis viruses in North America. Am J Trop Med Hyg 77 :58ā66.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 41 | 41 | 6 |
| Full Text Views | 424 | 139 | 0 |
| PDF Downloads | 247 | 71 | 0 |