Rapid polyvalent screening for largescale environmental Spiroplasma surveys

Abstract

Surface serology is an important determinant in Spiroplasma systematics. Reciprocal antigen/antibody reactions between spiroplasmas and individual antisera delineate the 38 described groups and species.However, reciprocal serology is impractical for large-scale studies. This report describes a successful, streamlined polyvalent screening approach used to examine isolates from an environmental survey.

Production and application of polyclonal and monoclonal antibodies against Spiroplasma eriocheiris

“…Deformation test was done according to the method reported before with slight modifications 33 . Briefly, 50 μl mAbs to be examined were added to the same volume of S. eriocheiris cultures in the logarithmic phase of growth.…”

An Australian environmental survey reveals moderate Spiroplasma biodiversity: characterization of four new serogroups and a continental variant

“…Log-phase cultures with helical morphology were used for serological analyses. Initially, cultures were screened using polyvalent screening mixtures (French et al 2009) and then in individual serological deformation (DF) assays (Williamson et al 1978). Antisera were produced against serologically unresolved isolates and added to initial screens and DF tests, as generated.…”

“…Antisera were produced against serologically unresolved isolates and added to initial screens and DF tests, as generated. New antisera produced for this study were prepared as described previously French et al 2009); animals were cared for in accordance with approved guidelines set forth in the ''Guide for the Care and Use of Laboratory Animals'' and their use was reviewed and approved by the Institutional Animal Care and Use Committee of Georgia Southern University. Anti-sera to established tabanid-associated serogroups and (or) subgroups were obtained from collections at Georgia Southern University (Statesboro, Georgia), the Agricultural Research Center (Beltsville, Maryland), and the National Institute of Allergy and Infectious Diseases Laboratory (Frederick, Maryland).…”

“…Overall serological analyses were completed as described previously (French et al 2009). Initial isolate screening was completed using polyvalent antisera mixtures in conjunction with successive rounds of dilution cloning (Whitcomb and Hackett 1987).…”