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A novel and simple colorimetric method for screening Giardia intestinalis and anti-giardial activity in vitro

Published online by Cambridge University Press:  01 September 1998

E. W. KANG
Affiliation:
School of Biochemistry and Molecular Genetics, University of New South Wales, Sydney, NSW 2052, Australia
K. CLINCH
Affiliation:
Industrial Research Limited, Gracefield Road, P.O. Box 31 310, Lower Hutt, New Zealand
R. H. FURNEAUX
Affiliation:
Industrial Research Limited, Gracefield Road, P.O. Box 31 310, Lower Hutt, New Zealand
J. E. HARVEY
Affiliation:
Industrial Research Limited, Gracefield Road, P.O. Box 31 310, Lower Hutt, New Zealand
P. J. SCHOFIELD
Affiliation:
School of Biochemistry and Molecular Genetics, University of New South Wales, Sydney, NSW 2052, Australia
A. M. GERO
Affiliation:
School of Biochemistry and Molecular Genetics, University of New South Wales, Sydney, NSW 2052, Australia

Abstract

A new and simple colorimetric method has been developed for determining activity in vitro against Giardia intestinalis. The microtitre plate assay is based upon the nucleoside hydrolase activity released from G. intestinalis by lysis. Action of the nucleoside hydrolase on the substrate analogue, 4-nitrophenyl β-d-ribofuranoside (NPR), gives rise to a coloured product which may be determined directly by the change in absorbance. A number of other such nucleoside analogues can be similarly used, but NPR is the preferred substrate, since it gives high enzymic activity at a relatively low substrate concentration. The IC50 values determined using this method for the known anti-giardials metronidazole, tinidazole and furazolidone were consistent with previously published values. The method is simple, does not involve radioisotopes or complex instrumentation, and thus provides a convenient method for screening potential anti-giardial agents.

Type
Research Article
Copyright
1998 Cambridge University Press

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