Development of an ELISA for Detection of Myxoma Virus-Specific Rabbit Antibodies: Test Evaluation for Diagnostic Applications on Vaccinated and Wild Rabbit Sera

Abstract

An enzyme-linked immunosorbent assay (ELISA) was developed and compared with 2 reference diagnostic tests (indirect immunofluorescence [IF] and complement fixation) to detect myxoma virus-specific antibodies in sera from 50 rabbits experimentally vaccinated with an attenuated strain of myxoma virus or with a Shope fibroma virus. The ELISA was highly specific (100% specificity) and sensitive (100%, 21 days after homologous vaccination). In a comparison of the ELISA with the IF test in 128 wild rabbits from France, discrepant results were obtained in only 11 (8.6%) animals, which were positive with the ELISA and negative with the IF test. The higher sensitivity and the good specificity of the ELISA was confirmed in a serologic survey of 118 rabbits from 2 Kerguelen (Indian Ocean) islands, where the prevalence of myxomatosis varied considerably. The ELISA is an alternative serologic test for diagnosis, vaccine evaluation, and seroepidemiologic surveys of myxomatosis.

References

Bradford

: 1976, A rapid and sensitive method for the quantification of micrograms quantities of protein utilizing the principle of protein-dye binding Anal Biochem 72:248–254

Chantal

Boucraut-Baralon

Ganière

: 1993, The complement fixation test on microtitration plates applied to the serology of myxomatosis. Comparison of the results with those of indirect immunofluorescence Rev Sci Tech Off Int Epizoot 12:895–907

Chantal

Gilbert

: 1996, Myxomatosis In: Manual of standards for diagnostic tests and vaccines, pp. 578–583. Office International des Epizooties, World Organization for Animal Health, Paris, France

Chapuis

Chantal

Bijlenga

: 1994, Myxomatosis without vectors on the subantarctic islands of Kerguelen three decades after its introduction CR Acad Sci Life Sci 317:174–182

Fenner

Marschall

Woodroofe

: 1953, Studies in the epidemiology of infectious myxomatosis of rabbits. 1. Recovery of Australian wild rabbit (Oryctolagus cuniculus) from myxomatosis under field conditions J Hyg. 51:225–244

Fenner

Myers

: 1978, Myxoma virus and myxomatosis in retrospect: the first quarter century of a new disease In: Viruses and environment, ed. Kurstok

Maramorosch

, 1st ed., pp. 539–570. Academic Press, New York, NY

Fenner

Ratcliff

: 1965, Myxomatosis in Oryctolagus cuniculus: the immunological response In: Myxomatosis, ed. Fenner

Ratcliff

, 1st ed., pp. 118–133. Cambridge University Press, London, England

Fenner

Woodroofe

: 1954, Protection of laboratory rabbits against myxomatosis by vaccination with Shope fibroma virus Aust J Exp Biol 32:653–668

Gardner

Hietala

Boyce

: 1996, Validity of using serological tests for diagnosis of diseases in wild animals Rev Sci Tech Off Int Epizoot 15:323–335

10.

Gilbert

Picavet

Chantal

: 1989, Diagnosis of myxomatosis: development of an indirect immunofluorescence technique Rev Sci Tech Off Int Epizoot 8:209–220

11.

Joubert

Duclos

Tuaillon

: 1982, Wild rabbits myxomatosis in South-eastern France Rev Med Vet 133:739–753

12.

Martin

Shoukri

Thorburn

: 1992, Evaluating the health status of herds based on tests applied to individuals Prev Vet Med 14:33–43

13.

McFadden

: 1988, Poxviruses in rabbits In: Virus diseases in laboratory and captive animals, ed. Darai

, 1st ed., pp. 37–62. Martinus Nijhoff, Boston, MA

14.

Rogan

Gladen

: 1978, Estimating prevalence from the results of a screening test Am J Epidemiol 107:71–76

15.

Saurat

Gilbert

Ganiere

: 1978, Etude d'une souche de virus myxomateux modifyé Rev Med Vet 129:415–451

16.

Shope

: 1938, Protection of rabbits against naturally acquired infectious myxomatosis by previous infection with fibroma virus Proc Soc Exp Biol 38:86–88

17.

Sobey

Conolly

Adams

: 1966, Myxomatosis. A simple method of sampling blood and testing for circulating soluble antigens or antibodies to them Aust J Sci 28:354–355

18.

Sobey

Conolly

Adams

: 1970, Myxomatosis: measuring antibodies to soluble antigens collected on filter paper strips Aust J Sci 32:331–332

19.

Thrusfield

: 1995, Diagnostic testing In: Veterinary epidemiology, ed. Thrusfield

, 2nd ed., pp. 266–285. Blackwell, Oxford, England

20.

Wetherall

Clay

King

: 1983, Humoral immunity to myxoma virus in wild rabbits Aust J Wild Res 10:277–285

21.

Woodroofe

Fenner

: 1965, Viruses of the myxoma-fibroma subgroup of the poxviruses. I. Plaque production in cultured cells, plaque reduction tests, and cross-protection tests in rabbits Aust J Exp Biol Med Sci 43:123–142