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    Alignment of the deduced amino acid sequence for 137 residues obtained from 14 strains of Sindbis-like virus and Whataroa virus described in Table 1. Dashes show amino acid insertions/deletions. Dots show sequence identity.

  • View in gallery

    Phylogenetic tree based on nucleotide sequences of the capsid gene of 19 Sindbis-like virus and Whataroa virus (WHAV) isolates. The tree was constructed using the neighbor-joining method. The tree was out-grouped by the sequence of the same region of Aura virus. Numbers indicate bootstrap confidence values. Vertical distances are arbitrary. Horizontal distances are proportional to the genetic distance as indicated by the bar. P/E = Paleoarctic-Ethiopian; SW = Southwest; O/A = Oriental-Australian.

  • 1

    Doherty RL, Bodey AS, Carew JS, 1969. Sindbis virus infection in Australia. Med J Aust 2 :1016–1017.

  • 2

    Guard RW, McAuliffe MJ, Stallman ND, Bramton BA, 1982. Haemorrhagic manifestations with Sindbis infection. Case Report. Pathology 14 :89–90.

    • Search Google Scholar
    • Export Citation
  • 3

    Rentier-Delrue F, Young NA, 1980. Genomic divergence among Sindbis virus strains. Virology 106 :59–70.

  • 4

    Olson K, Trent DW, 1985. Genetic and antigenic variations among geographical isolates of Sindbis virus. J Gen Virol 66 :797–810.

  • 5

    Sammels LM, Lindsay MD, Poidinger M, Coelen JR, Mackenzie JS, 1999. Geographic distribution and evolution of Sindbis virus in Australia. J Gen Virol 80 :739–748.

    • Search Google Scholar
    • Export Citation
  • 6

    Saleh SM, Poidinger M, Mackenzie JS, Broom AK, Lindsay MD, Hall RA. Complete genomic sequence of the Australian southwest genotype of Sindbis virus: comparisons with other Sindbis strains and identification of a unique deletion in the 3′-untranslated region. Virus Genes 26 :317–327.

    • Search Google Scholar
    • Export Citation
  • 7

    Powers AM, Brault AC, Shirako Y, Strauss EG, Kang W, Strauss JH, Weaver SC, 2001. Evolutionary relationships and systematics of the alphaviruses. J Virol 75 :10118–10131.

    • Search Google Scholar
    • Export Citation
  • 8

    Liang GD, Li L, Zhou GL, Fu SH, Li QP, Li FS, He HH, Jin Q, He Y, Chen BQ, Hou YD, 2000. Isolation and complete nucleotide sequence of a Chinese Sindbis-like virus. J Gen Virol 81 :1347–1351.

    • Search Google Scholar
    • Export Citation
  • 9

    Poidinger M, Roy S, Hall RA, Turley PJ, Scherret JH, Lindsay MD, Broom AK, Mackenzie JS, 1997. Genetic stability among temporally and geographically diverse isolates of Barmah Forest virus. Am J Trop Med Hyg 572 :230–234.

    • Search Google Scholar
    • Export Citation
  • 10

    Adams SC, Broom AK, Sammels LM, Hartnett AC, Howard MJ, Coelen RJ, Mackenzie JS, Hall RA, 1995. Glycosylation and antigenic variation among Kunjin virus isolates. Virology 206 :49–56.

    • Search Google Scholar
    • Export Citation
  • 11

    Hall RA, Kay BH, Burgess GW, Clancy P, Fanning ID, 1990. Epitope analysis of the envelope and non-structural glycoproteins of Murray Valley encephalitis virus. J Gen Virol 71 :2923–2930.

    • Search Google Scholar
    • Export Citation
  • 12

    Burgess GW, Clancy P, Yan M, 1993. Development of monoclonal antibody based diagnostics for alphaviruses. Arbovirus Res Aust 6 :87–89.

  • 13

    Stanley J, Cooper SJ, Griffin DE, 1985. Alphavirus neurovirulence: monoclonal antibodies discriminating wild-type from neuroadapted Sindbis virus. J Virol 56 :110–119.

    • Search Google Scholar
    • Export Citation
  • 14

    Sellner LN, Coelen RJ, Mackenzie JS, 1992. A one-tube, one manipulation RT-PCR reaction for detection of Ross River virus. J Virol Methods 40 :255–264.

    • Search Google Scholar
    • Export Citation
  • 15

    Strauss EG, Rice CM, Strauss JH, 1984. Complete nucleotide sequence of the genomic RNA of Sindbis virus. Virology 133 :92–110.

  • 16

    Thompson JD, Higgins DG, Gibson TJ, 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22 :4673–4680.

    • Search Google Scholar
    • Export Citation
  • 17

    Felsenstein J, 1995. PHYLIP Manual, Version 3.57c. Seattle: University of Washington.

  • 18

    Russell RC, Cloonan MJ, Doggett SL, Clancy J, Haniotis J,Wells P, Fennell M, Cunningham A, Hueston L, Marchetti M, 1997. Surveillance of arboviruses and vectors in NSW, 1993–1996. Arbovirus Res Aust 7 :228–234.

    • Search Google Scholar
    • Export Citation
  • 19

    Calisher CH, Karabatsos N, Lazuick JS, Monath TP, Wolff KL, 1988. Reevaluation of the western equine encephalitis antigenic complex of alphaviruses (family Togaviridae) as determined by neutralization tests. Am J Trop Med Hyg 38 :447–452.

    • Search Google Scholar
    • Export Citation
  • 20

    Shirako Y, Niklasson B, Dalrymple JM, Strauss EG, Strauss GH, 1991. Structure of the Ockelbo genome and its relationship to other Sindbis viruses. Virology 182 :753–764.

    • Search Google Scholar
    • Export Citation
  • 21

    Simpson DA, Davis ND, Lin S, Russel D, Johnston RE, 1996. Complete nucleotide sequence and full-length cDNA clone of S.A.A.R. 86, a South African alphavirus related to Sindbis. Virology 222 :464–469.

    • Search Google Scholar
    • Export Citation
  • 22

    Maguire T, Miles JA, Casals J, 1967. Whataroa virus, a group A arbovirus isolated in South Westland, New Zealand. Am J Trop Med Hyg 16 :371–373.

    • Search Google Scholar
    • Export Citation
  • 23

    Miles JAR, 1973. The ecology of Whataroa virus, an alphavirus, in south Westland, New Zealand. J Hyg 71 :701–713.

 

 

 

 

ANTIGENIC AND GENETIC TYPING OF WHATAROA VIRUSES IN AUSTRALIA

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  • 1 Department of Microbiology and Parasitology, University of Queensland, Brisbane, Queensland Australia; Department of Microbiology, University of Western Australia, Perth, Western Australia, Australia; Western Australian Health Department, Perth, Western Australia, Australia; Department of Medical Entomology, Institute of Clinical Pathology and Medical Research, University of Sydney, Westmead Hospital, Westmead, New South Wales, Australia; Department of Microbiology, Prince Henry Hospital, Little Bay, New South Wales, Australia; Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado; Department of Microbiology and Immunology, James Cook University, Townsville, Queensland, Australia

We recently characterized three novel alphaviruses isolated from mosquitoes captured in New South Wales, Australia. Initial cross-neutralization studies revealed antigenic similarity to the Sindbis virus (SINV)–like Whataroa virus (WHAV), heretofore found only in New Zealand. Nucleotide sequence analysis showed that the WHAV-like viruses shared >99% nucleotide sequence similarity with each other, and 96–97% similarity with prototype WHAV. Enzyme-linked immunosorbent assay reactions of a panel of monoclonal antibodies to SINV showed that the novel WHAV-like viruses displayed identical binding patterns and were antigenically distinct from all SINV isolates examined. Although these viruses displayed a similar binding pattern to prototype WHAV, three monoclonal antibodies discriminated them from the New Zealand virus. Our results suggest that these novel alphaviruses are antigenic variants of WHAV and represent the first reported isolations of this virus from outside New Zealand. The monoclonal antibodies used in this study will be useful for typing new SINV and SINV-like isolates.

INTRODUCTION

Sindbis virus (SINV) is a mosquito-transmitted alphavirus (family Togaviridae, genus Alphavirus) found in Europe, Africa, Asia, and Australasia and has been responsible for outbreaks of fever and rash.1,2 Previous studies have demonstrated the existence of two major genotypes of SINV, the Paleoarctic-Ethiopian (P/E) genotype found in Europe and Africa, and the Oriental-Australian (O/A) genotype found in Asia and Australasia.3,4 More recent genetic studies of SINV isolates from mosquitoes trapped in the southwest region of Western Australia between 1990 and 1998 have demonstrated the existence of a third genetic lineage of SINV, the southwest (SW) genotype.5,6 Close antigenic relatives of SINV (SINV-like viruses) also have been isolated from Europe (Ockelbo virus), Azerbaijan (Kyzylagach virus), China (strain XJ160), South America (Aura virus), and Whataroa virus (WHAV) from New Zealand.7,8 On the basis of genetic differences, the latter two are considered species distinct from that of SINV.

Here we report the identification and characterization of SINV-like viruses from mosquitoes trapped between 1989 and 1990 in New South Wales, Australia. We also describe the use of a panel of monoclonal antibodies (MAbs) prepared to three SINV genotypes for precise typing of SINV and SINV-like viruses.

MATERIALS AND METHODS

Cell cultures.

African Green monkey kidney (Vero) cells were grown in medium M199 (Gibco-BRL, Gaithersburg, MD) supplemented with 10 mM HEPES, antibiotics (100 mg/L of benzylpenicillin and 10g/L of gentamicin), and 10% fetal bovine serum (FBS) for growth, 2% for maintenance, and incubated at 37°C. A line of Aedes albopictus (C6/36) cells was cultured in medium M199 with no HEPES and supplemented with FBS as above for growth or maintenance. The cells were incubated at 28°C in a humidified atmosphere in the presence of 5% CO2.

Viruses.

Details of the SINV isolates used in this study are listed in Table 1. Stocks of each virus were prepared in cell cultures by inoculating at a multiplicity of infection of one. Virions in the culture supernatant were concentrated by polyethylene glycol precipitation, purified by sucrose gradient centrifugation, and the virion RNA was extracted as previously described.5,9

Monoclonal antibodies.

Ten MAbs were prepared to SINV: three to the SW genotype strain SW6562 and seven to the 33130 strain from the O/A genotype using methods described previously.10 The isotype, protein specificity, and biologic activity of these MAbs were determined as previously described,11 and are listed in Table 2. Reference MAbs 11F4 and 2F2 were generously provided by TropBio Pty Ltd. (Townsville, Queensland, Australia). Monoclonal antibody 11F4 was previously shown to be alphavirus group-specific,12 while MAb 2F2, raised to the Australian prototype strain MRM39, was shown to be SINV-specific (Clancy P, 1991. Production and Application of Monoclonal Antibodies to the Australian Alphaviruses. MSc Thesis. School of Tropical Veterinary Science and Agriculture, James Cook University of North Queensland, Townsville, Queensland, Australia). Four additional reference MAbs (161, 2, 129, and 17C), raised to SINV prototype strain AR339,13 were a gift from Dianne Griffin (Johns Hopkins Bloomberg School of Public Health, Baltimore, MD). The reaction of each MAb with each SINV and SINV-like strain in this study was determined by enzyme-linked immunosorbent assay, as previously described.10

Polymerase chain reaction and sequencing.

A single-tube reverse transcriptase–polymerase chain reaction (RT-PCR) procedure was performed on each SINV isolate using a protocol based on a previous study,14 using oligonucleotides designed from regions of SINV sequences taken from GenBank.15 Primer pairs were designed from each region (backward and forward primers) to amplify a 459 nucleotide region of the capsid gene. Specific details about oligonucleotide primers used to generate the sequence have been previously described.6 All sequencing of the PCR products was performed using the PRISM Dye Terminator Sequencing Kit (Applied Biosystems, Foster City, CA) and analyzed on a 373A automated sequencer (Applied Biosystems).

The consensus sequences were then aligned using the program Clustal W16 and phylogenetic data were generated using some of the programs in the PHYLIP package.17 Bootstrap random sampling was used to place confidence values on grouping within trees. The GenBank accession numbers for the sequences reported in this paper are AF035087 and AY22742-AY22744.

RESULTS

Initial identification of WHAV-like viruses by cross-neutralization.

Several virus isolates were obtained from mosquitoes trapped in southern New South Wales between 1989 and 1990 using previously described methods.18 These viruses showed similar characteristics in suckling mice and cell cultures and were initially identified as alphaviruses by an indirect immunofluorescence assay using an extensive panel of arbovirus polyclonal typing reagents supplied by the U.S. National Institutes of Health (Bethesda, MD). Further typing of three of these viruses (1209, 1337, 1264) was performed by cross-neutralization with a panel of reference alphaviruses using previously described methods.19 Two-way cross neutralization of each test virus with WHAV provided a tentative identification as WHAV-like viruses (Table 3).

Confirmation of WHAV isolates by partial genomic sequencing.

The sequences for 459 nucleotides in the capsid gene of the three WHAV-like virus isolates and the prototype WHAV (M78 strain) were obtained from viral RNA by RT-PCR and automated sequencing. These sequences were aligned and compared with corresponding sequences previously published for SINV strains from the P/E and O/A geographic regions (Sammels LM, 1995. Molecular Epidemiology of Ross River and Sindbis Viruses. PhD Thesis. The University of Western Australia, Nedlands, Perth, Western Australia).15,20,21 The nucleotide sequences of the four SINV strains from the SW genotype that we had previously obtained6 were also used in the alignments of the capsid gene.

Alignment of the nucleotide sequences in the capsid gene confirmed that the WHAV-like strains 1209, 1264, and 1337 were virtually identical to one another (>99% identity) and most closely related to the M78 strain of WHAV (>96% identity). These four viruses also shared a number of amino acid substitutions and two amino acid insertions in the capsid protein compared with prototype SINV (Figure 1). In contrast, there was significant nucleotide divergence between the three WHAV-like strains and the P/E (67–69% identity), O/A (66–67% identity), and SW (63–64% identity) isolates. The phylogenic tree constructed from the nucleotide sequence data obtained from the capsid gene (Figure 2) defined four clusters of SINV and WHAV-like viruses. Three corresponded to the P/E, O/A, and SW SINV genotypes. The fourth group (WHAV-like) included the prototype New Zealand WHAV strain M78 and the three WHAV-like strains isolated in New South Wales. Bootstrap analysis of nucleotide sequence data separated the viruses into the P/E, O/A, SW, and WHAV-like groups in all 100 randomly generated tests. Clustering of the three WHAV-like viruses with the M78 strain of WHAV occurred with 100% confidence. These results confirm the identity of isolates 1209, 1264, and 1337 as new strains of WHAV.

Typing of SINV and WHA isolates using MAbs.

A panel of SINV-reactive MAbs, including 10 MAbs produced for this study, was used to investigate the antigenic variation of SINV and WHAV isolates. From the binding pattern of these MAbs to 16 isolates of SINV and WHAV isolates cultured in C6/36 cells, five antigenic groups were identified: P/E SINV, O/A SINV, SW SINV, the New Zealand WHAV strain M78, and the Australian WHAV isolates (Table 4). The three MAbs raised to the SW isolate 6562 (9A10, 3G7, and 1C8) recognized the four SW SINV isolates, but showed different patterns of reactivity with the rest of the strains examined. Monoclonal antibody 9A10 was specific for the SW genotype and failed to recognize any other isolates used in the study. Monoclonal antibody 3G7 also recognized the Egyptian SINV prototype AR339 strain. Monoclonal antibody 1C8 reacted with strain AR339 and with the four WHAV strains (M78, 1209, 1264, and 1337). The seven MAbs raised to O/A isolate 33130 (3A7, 3F9, 1C7, 2A9, 2G11, 4F3, and 5H9) reacted with all O/A isolates used in this study, including strain MRE16, isolated from Malaysia. Monoclonal antibody 3A7 was specific for the O/A genotype and failed to recognize any other SINV isolate. Monoclonal antibodies 3F9 and 4F3 also reacted with strain AR339 and the WHAV isolates. Monoclonal antibodies 1C7, 2G11, and 5H9 recognized all isolates of SINV and WHAV. Monoclonal antibody 2A9 recognized all SINV and WHAV strains with the exception of WHAV strain M78. All SINV strains used in this study were recognized by the alphavirus-specific, reference MAb 11F4 prepared to Barmah Forest virus and the SINV-specific reference MAb 2F2 (prepared to strain MRM39). However, MAb 2F2 failed to recognize the four WHAV strains. Monoclonal antibodies 161, 2, 17C, and 129, which were raised to the SINV prototype strain AR339,13 exhibited variable cross-reactivity with the SINV isolates examined in this study. Monoclonal antibody 17C recognized all SINV and WHAV isolates. Monoclonal antibody 161 only bound to strain AR339 and the four WHAV isolates. Monoclonal antibody 2 recognized all isolates except WHAV strain M78. Monoclonal antibody 129 failed to recognize the SW SINV isolates and WHAV strain M78.

DISCUSSION

The three novel alphavirus isolates at the focus of this study were shown to share a high level of nucleotide sequence homology with each other (>99% identity) and with prototype WHAV (approximately 96% identity), consistent with their identification as a newly recognized strain of WHAV. These are the first Australian isolates of WHAV, which was originally isolated in 1962 in south Westland of New Zealand, near the township of Whataroa, and is thought to be maintained in mosquito-bird transmission cycles.22,23

The MAb binding studies also demonstrated antigenic similarity between prototype WHAV (strain M78) and the Australian WHAV isolates. However, two MAbs raised to the E2 protein of P/E SINV strain AR339 (129 and 2) and one to the E2 protein of O/A SINV strain 33130 (2A9) differentiated the three Australian WHAV isolates from the prototype virus, suggesting the former are antigenic variants of WHAV. The MAb binding patterns to the remaining SINV strains used in this study placed the viruses into three antigenic groups that correlated precisely with the genotypes identified by sequence comparisons. These MAbs will be valuable tools for the rapid identification and subtyping of new SINV and SINV-like isolates. A typing panel that included MAbs 2F2, 5H9, 3G7, 9A10, 2A9, 1C8, and 3A7 would be sufficient for this purpose.

The isolation of antigenic variants of WHAV from mosquitoes in New South Wales, approximately 3,000 km west of New Zealand, suggests the virus was introduced to Australia from New Zealand, possibly by migrating birds, and has evolved independently in the new habitat. The possibility that the virus was introduced from Australia to New Zealand prior to 1962 is a less likely scenario because regular arbovirus surveillance programs in Australia over several decades have found no evidence of the virus prior to 1989 or since 1990.

Three distinct genetic lineages of SINV or WHAV have now been shown to occur in Australia. One of these, the O/A SINV genotype, circulates in bird-mosquito transmission cycles throughout the continent as well as in Papua New Guinea, and Sarawak, Malaysia. In contrast, the other two lineages (the SW SINV genotype and the new WHAV strain) appear to be restricted to discrete geographic areas located in southwestern Australia and southern New South Wales, respectively. To understand the ecology and epidemiology of these viruses, including the identification of their vertebrate hosts and arthropod vectors, further investigations are required.

Table 1

Details of the virus isolates used in this study

Isolate nameLocationDate collectedSource
* Prototype Sindbis virus.
† These mosquito species were previous found in the genus Aedes.
AR339*Cairo, Egypt1953Culex univittatus
SAAR86South Africa1954Mixed pool Culex spp.
Edsbyn82Sweden1982Mixed pool Culiseta spp.
MRE16Sarawak, Malaysia1975Unknown
MRM39Kowanyama, north Queensland (QLD)1960Cx. annulirostris
MK6962Joinjakaka, Papua New Guinea1966Ficalbia flavens
RRD800Ross River Dam, north QLD1991Cx. annulirostris
WK465Camballin, West Kimberley, Western Australia (WA)1979Cx. annulirostris
K3028Nullagine, Pilbara, WA1990Ochlerotatus pseudonormanensis
K12710Billiluna, South Kimberley, WA1993Cx. annulirostris
33130Mandurah, WA1993Oc. camptorhynchus
CY174Pajinka, QLD1996Cx. annulirostris
SW6562Southern suburbs, Perth, southwest WA1990Cx. annulirostris
SW20055Peel region, southwest WA1992Cx. annulirostris
SW25713Leschenault region, southwest, WA1992Oc. camptorhynchus
SW33029Peel region, southwest WA1993Oc. camptorhynchus
M78New Zealand1962Cx. pervigilans
1209Griffith, New South Wales (NSW)1990Cx. annulirostris
1264Griffith, NSW1989Cx. annulirostris
1337Leeton, NSW1989Cx. annulirostris and Anopheles annulipes
Table 2

Properties of monoclonal antibodies used in this study*

Virus strain†Monoclonal antibodyIsotypeReactive proteinNeutralizing activityHI activity
* HI = hemagglutination inhibition; E = envelope; NT = not tested; C = capsid.
† Virus strain used for monoclonal antibody production.
BFV BH219311F4IgG1E1/E2NT+
SINV P/E AR339161IgG2bE1++
129IgG3E2++
2IgG3E2+
17CIgG2aE1++
SINV O/A MRM392F2IgG1E1/E2NTNT
SINV O/A 331303A7IgG2aE2++
4F3IgME2+
5H9IgG2bE1+
1C7IgG2aE1+
2G11IgG1E2
2A9IgG2aE2++
3F9IgG2aC
SINV SW 65621C8IgMC
3G7IgG2aC+
9A10IgG1E2
Table 3

Cross-neutralization titers of Sindbis virus (SINV)–like isolates with reference alphaviruses*

Antiserum to
Virus120912641337SINVSAGVWHAV
* There was no reaction of SINV-like isolates in either direction with Semliki Forest virus (homologous antibody titer-160), Bebaru virus (40), Getah virus (320), Barmah Forest virus, (320) or Ross River virus (40). SINV = prototype Sindbis virus; SAGV = prototype Sagiyama virus; WHAV = prototype Whataroa virus. The neutralization titer is shown as the reciprocal of the highest serum dilution to inhibit plaque numbers by 90%. Underlined values are homologous titers. – = <20. Bold values indicate the two-way cross-neutralization between WHAV and the three test viruses.
12098040
1264801601608020
133716016032032040
SINV202032040
SAGV20640
WHAV204016040
Table 4

Reactivity of SINV monoclonal antibodies (MAbs) with 15 strains of SINV and WHAV in an ELISA*

Viruses
P/EO/ASWWHAV
MAbAR339MRM39MRE16CY17433130K12710K30286562200552571333029M78120912641337
* SINV = prototype Sindbis virus; WHAV = prototype Whataroa virus; ELISA = enzyme-linked immunosorbent assay.
† Alphavirus-specific MAb.12
‡ MAb raised against SINV prototype AR339.13
§ MAb raised against Australian SINV prototype MRM39.
¶ MAb raised against O/A SINV strain 33130.
# MAb raised against SW SINV strain 6562.
11F4†+++++++++++++++
161‡+++++
129‡++++++++++
2‡++++++++++++++
17C‡+++++++++++++++
2F2§+++++++++++
4F3¶+++++++++++
3A7¶++++++
5H9¶+++++++++++++++
1C7¶+++++++++++++++
2G11¶+++++++++++++++
3F9¶+++++++++++
2A9¶++++++++++++++
1C8#+++++++++
3G7#+++++
9A10#++++
Figure 1.
Figure 1.

Alignment of the deduced amino acid sequence for 137 residues obtained from 14 strains of Sindbis-like virus and Whataroa virus described in Table 1. Dashes show amino acid insertions/deletions. Dots show sequence identity.

Citation: The American Journal of Tropical Medicine and Hygiene Am J Trop Med Hyg 71, 3; 10.4269/ajtmh.2004.71.262

Figure 2.
Figure 2.

Phylogenetic tree based on nucleotide sequences of the capsid gene of 19 Sindbis-like virus and Whataroa virus (WHAV) isolates. The tree was constructed using the neighbor-joining method. The tree was out-grouped by the sequence of the same region of Aura virus. Numbers indicate bootstrap confidence values. Vertical distances are arbitrary. Horizontal distances are proportional to the genetic distance as indicated by the bar. P/E = Paleoarctic-Ethiopian; SW = Southwest; O/A = Oriental-Australian.

Citation: The American Journal of Tropical Medicine and Hygiene Am J Trop Med Hyg 71, 3; 10.4269/ajtmh.2004.71.262

Authors’ addresses: Suha M. Saleh, Michael Poidinger, John S. Mackenzie, and Roy A. Hall, Department of Microbiology and Parasitology, School of Molecular and Microbial Sciences, University of Queensland, Brisbane 4072, Australia, Fax: 61-7-3365-4620, E-mail: roy.hall@mailbox.uq.edu.au. Annette K. Broom, Department of Microbiology, The University of Western Australia, Perth, Western Australia 6009, Australia. Michael D. A. Lindsay, Mosquito-Borne Disease Control Branch, Western Australian Department of Health, Perth, Western Australia 6849, Australia. Richard C. Russell, Department of Medical Entomology, Institute of Clinical Pathology and Medical Research, University of Sydney, Westmead Hospital, Westmead, New South Wales 2145, Australia. Michael J. Cloonan, Department of Microbiology, Prince Henry Hospital, Little Bay, New South Wales 2036, Australia. Charles H Calisher, Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523. Graham W. Burgess, Microbiology and Immunology, James Cook University, Townsville, Queensland, 4811, Australia.

Acknowledgments: We dedicate this work to the memory of Dr. Robert E. Shope. Dr. Shope assisted us in this work, as he did for so many of our efforts, by providing polyclonal alphavirus typing reagents. We also thank Dianne Griffin for providing reference monoclonal antibodies, Danielle Heilpern, Peter Wells, John Haniotis, and Stephen Doggett for laboratory assistance in virus isolation, and Robert Huestis for assistance with sequence submission to GenBank.

Financial support: This study was supported by grants from the National Health and Medical Research Council, Australia.

REFERENCES

  • 1

    Doherty RL, Bodey AS, Carew JS, 1969. Sindbis virus infection in Australia. Med J Aust 2 :1016–1017.

  • 2

    Guard RW, McAuliffe MJ, Stallman ND, Bramton BA, 1982. Haemorrhagic manifestations with Sindbis infection. Case Report. Pathology 14 :89–90.

    • Search Google Scholar
    • Export Citation
  • 3

    Rentier-Delrue F, Young NA, 1980. Genomic divergence among Sindbis virus strains. Virology 106 :59–70.

  • 4

    Olson K, Trent DW, 1985. Genetic and antigenic variations among geographical isolates of Sindbis virus. J Gen Virol 66 :797–810.

  • 5

    Sammels LM, Lindsay MD, Poidinger M, Coelen JR, Mackenzie JS, 1999. Geographic distribution and evolution of Sindbis virus in Australia. J Gen Virol 80 :739–748.

    • Search Google Scholar
    • Export Citation
  • 6

    Saleh SM, Poidinger M, Mackenzie JS, Broom AK, Lindsay MD, Hall RA. Complete genomic sequence of the Australian southwest genotype of Sindbis virus: comparisons with other Sindbis strains and identification of a unique deletion in the 3′-untranslated region. Virus Genes 26 :317–327.

    • Search Google Scholar
    • Export Citation
  • 7

    Powers AM, Brault AC, Shirako Y, Strauss EG, Kang W, Strauss JH, Weaver SC, 2001. Evolutionary relationships and systematics of the alphaviruses. J Virol 75 :10118–10131.

    • Search Google Scholar
    • Export Citation
  • 8

    Liang GD, Li L, Zhou GL, Fu SH, Li QP, Li FS, He HH, Jin Q, He Y, Chen BQ, Hou YD, 2000. Isolation and complete nucleotide sequence of a Chinese Sindbis-like virus. J Gen Virol 81 :1347–1351.

    • Search Google Scholar
    • Export Citation
  • 9

    Poidinger M, Roy S, Hall RA, Turley PJ, Scherret JH, Lindsay MD, Broom AK, Mackenzie JS, 1997. Genetic stability among temporally and geographically diverse isolates of Barmah Forest virus. Am J Trop Med Hyg 572 :230–234.

    • Search Google Scholar
    • Export Citation
  • 10

    Adams SC, Broom AK, Sammels LM, Hartnett AC, Howard MJ, Coelen RJ, Mackenzie JS, Hall RA, 1995. Glycosylation and antigenic variation among Kunjin virus isolates. Virology 206 :49–56.

    • Search Google Scholar
    • Export Citation
  • 11

    Hall RA, Kay BH, Burgess GW, Clancy P, Fanning ID, 1990. Epitope analysis of the envelope and non-structural glycoproteins of Murray Valley encephalitis virus. J Gen Virol 71 :2923–2930.

    • Search Google Scholar
    • Export Citation
  • 12

    Burgess GW, Clancy P, Yan M, 1993. Development of monoclonal antibody based diagnostics for alphaviruses. Arbovirus Res Aust 6 :87–89.

  • 13

    Stanley J, Cooper SJ, Griffin DE, 1985. Alphavirus neurovirulence: monoclonal antibodies discriminating wild-type from neuroadapted Sindbis virus. J Virol 56 :110–119.

    • Search Google Scholar
    • Export Citation
  • 14

    Sellner LN, Coelen RJ, Mackenzie JS, 1992. A one-tube, one manipulation RT-PCR reaction for detection of Ross River virus. J Virol Methods 40 :255–264.

    • Search Google Scholar
    • Export Citation
  • 15

    Strauss EG, Rice CM, Strauss JH, 1984. Complete nucleotide sequence of the genomic RNA of Sindbis virus. Virology 133 :92–110.

  • 16

    Thompson JD, Higgins DG, Gibson TJ, 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22 :4673–4680.

    • Search Google Scholar
    • Export Citation
  • 17

    Felsenstein J, 1995. PHYLIP Manual, Version 3.57c. Seattle: University of Washington.

  • 18

    Russell RC, Cloonan MJ, Doggett SL, Clancy J, Haniotis J,Wells P, Fennell M, Cunningham A, Hueston L, Marchetti M, 1997. Surveillance of arboviruses and vectors in NSW, 1993–1996. Arbovirus Res Aust 7 :228–234.

    • Search Google Scholar
    • Export Citation
  • 19

    Calisher CH, Karabatsos N, Lazuick JS, Monath TP, Wolff KL, 1988. Reevaluation of the western equine encephalitis antigenic complex of alphaviruses (family Togaviridae) as determined by neutralization tests. Am J Trop Med Hyg 38 :447–452.

    • Search Google Scholar
    • Export Citation
  • 20

    Shirako Y, Niklasson B, Dalrymple JM, Strauss EG, Strauss GH, 1991. Structure of the Ockelbo genome and its relationship to other Sindbis viruses. Virology 182 :753–764.

    • Search Google Scholar
    • Export Citation
  • 21

    Simpson DA, Davis ND, Lin S, Russel D, Johnston RE, 1996. Complete nucleotide sequence and full-length cDNA clone of S.A.A.R. 86, a South African alphavirus related to Sindbis. Virology 222 :464–469.

    • Search Google Scholar
    • Export Citation
  • 22

    Maguire T, Miles JA, Casals J, 1967. Whataroa virus, a group A arbovirus isolated in South Westland, New Zealand. Am J Trop Med Hyg 16 :371–373.

    • Search Google Scholar
    • Export Citation
  • 23

    Miles JAR, 1973. The ecology of Whataroa virus, an alphavirus, in south Westland, New Zealand. J Hyg 71 :701–713.

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