Specific enzyme immunoassays for the rapid detection of Ross River virus in cell cultures inoculated with infected mosquito homogenates

Oliveira, Nidia M M; Broom, A.K.; Lindsay, M.D.; Mackenzie, John S.; Kay, Brian H.; Hall, Roy A.

doi:10.1016/0928-0197(94)00067-5

Cited by 9 publications

(5 citation statements)

References 19 publications

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“…At 10 days postinfection, the mosquitoes were anaesthetized, and the bodies and legs of individuals were separated. Leg samples were tested for RRV using a cell culture enzyme‐linked immunosorbent assay (Oliveira et al., ) at QIMR Berghofer, Queensland. Of the 125 Ae.…”

Section: Methodsmentioning

confidence: 99%

Effective mosquito and arbovirus surveillance using metabarcoding

Batovska

Lynch

Cogan

et al. 2017

Molecular Ecology Resources

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Effective vector and arbovirus surveillance requires timely and accurate screening techniques that can be easily upscaled. Next‐generation sequencing (NGS) is a high‐throughput technology that has the potential to modernize vector surveillance. When combined with DNA barcoding, it is termed ‘metabarcoding.’ The aim of our study was to establish a metabarcoding protocol to characterize pools of mosquitoes and screen them for virus. Pools contained 100 morphologically identified individuals, including one Ross River virus (RRV) infected mosquito, with three species present at different proportions: 1, 5, 94%. Nucleic acid extracted from both crude homogenate and supernatant was used to amplify a 269‐bp section of the mitochondrial cytochrome c oxidase subunit I (COI) locus. Additionally, a 67‐bp region of the RRV E2 gene was amplified from synthesized cDNA to screen for RRV. Amplicon sequencing was performed using an Illumina MiSeq, and bioinformatic analysis was performed using a DNA barcode database of Victorian mosquitoes. Metabarcoding successfully detected all mosquito species and RRV in every positive sample tested. The limits of species detection were also examined by screening a pool of 1000 individuals, successfully identifying the species and RRV from a single mosquito. The primers used for amplification, number of PCR cycles and total number of individuals present all have effects on the quantification of species in mixed bulk samples. Based on the results, a number of recommendations for future metabarcoding studies are presented. Overall, metabarcoding shows great promise for providing a new alternative approach to screening large insect surveillance trap catches.

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Section: Methodsmentioning

confidence: 99%

Effective mosquito and arbovirus surveillance using metabarcoding

Batovska

Lynch

Cogan

et al. 2017

Molecular Ecology Resources

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“…The mosquitoes were homogenized using sterile micropestles and a cordless motor (Quantum ScientiÞc, Brisbane, Australia) and centrifuged at 8,000 ϫ g for 10 min at 4ЊC. Supernatants were tested for RR and Barmah Forest (BF) viruses by Aedes albopictus (Skuse) salivary gland (C6/36) cell culture enzyme immunoassay (Oliveira et al 1995). Conßuent monolayers of C6/36 cells in 96-well tissue culture plates (Corning, Corning, NY) were inoculated with mosquito homogenates and incubated at 28ЊC with 5% carbon dioxide for 4 d. Fixed cell monolayers were examined for RR and BF antigens using RR speciÞc (3B2), BF speciÞc (3B3), and alphavirus (RR, BF, Getah, Semliki Forest, and Chikungunya viruses) cross reactive (B10) monoclonal antibodies (Mabs) (Kay et al 1996, Ritchie et al 1997.…”

Section: Methodsmentioning

confidence: 99%

Definition of Ross River Virus Vectors at Maroochy Shire, Australia

Ryan

Kay

2000

J Med Entomol

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Evidence of Ross River (RR) virus infection in field-collected mosquitoes and data from laboratory vector competence experiments incriminated a range of mosquito species as important vectors of RR virus in Maroochy Shire, Queensland, Australia. Nine RR and 2 Barmah Forest virus isolates were recovered from 27,529 mosquitoes collected in light traps from Maroochy Shire during 1996. Nine of the 10 most abundant mosquito species collected in light traps were fed on blood containing the B94/20 strain RR isolated from Queensland in 1994. All species except for Culex sitiens Wiedemann were susceptible to experimental infection. Evidence of RR virus transmission to mice was found with Aedes vigilax (Skuse), Aedes funereus (Theobald), Aedes procax (Skuse), Culex annulirostris Skuse, Mansonia uniformis (Theobald) and Culex australicus Dobrotworsky & Drummond. Aedes notoscriptus (Skuse) and Aedes multiplex (Theobald) were susceptible to RR virus infection, although there was no evidence of virus transmission. Based on adult abundance and vector competence results, freshwater species such as Cx. annulirostris, Ae. procax, and Ae. funereus, and saltmarsh Ae. vigilax, appear to be important vectors of RR virus in Maroochy Shire and control programs should be revised to include these species.

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“…As less virus was likely to be present in saliva samples compared with body or leg samples, a cell‐culture enzyme linked immunosorbent assay (ELISA), based on methods described by Oliveira et al . (1995), was evaluated for testing the contents of the capillary tubes.…”

Section: Methodsmentioning

confidence: 99%

Vector competence of Coquillettidia linealis (Skuse) (Diptera: Culicidae) for Ross River and Barmah Forest viruses

Jeffery

Ryan

Lyons

et al. 2002

Australian Journal of Entomology

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Coquillettidia linealis is a severe pest on some of the Moreton Bay islands in Queensland, Australia, but little is known of its breeding habitats and biology. Because of its high abundance and its association with Ross River (RR) and Barmah Forest (BF) viruses by field isolation, its vector competence was evaluated in the laboratory by feeding dilutions of both viruses in blood. For RR, Cq. linealis was of comparable efficiency to Ochlerotatus vigilax (Skuse), recognised as being a major vector. Results were as follows for Cq. linealis and Oc. vigilax , respectively: dose to infect 50%, 10 2.2 and <10 1.7 CCID 50 / mosquito; 88% and 90% disseminated infection at 4 days postinfection; transmission at 4 days with rates of 68-92% and 25-60%. For BF dose to infect 50%, 10 2.7 and 10 2.0 ; disseminated infection rates on first transmission day (day 6), 40% and 70%; transmission rates of 8-16% and 0-10%. As a capillary-tube method was used rather than suckling mice to demonstrate transmission, transmission rates may be underestimates. This, the first study of the vector competence of Cq. linealis in Australia, demonstrates that this species deserves control on the southern Moreton Bay islands.

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Specific enzyme immunoassays for the rapid detection of Ross River virus in cell cultures inoculated with infected mosquito homogenates

Cited by 9 publications

References 19 publications

Effective mosquito and arbovirus surveillance using metabarcoding

Effective mosquito and arbovirus surveillance using metabarcoding

Definition of Ross River Virus Vectors at Maroochy Shire, Australia

Vector competence of Coquillettidia linealis (Skuse) (Diptera: Culicidae) for Ross River and Barmah Forest viruses

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