Detection of viable Giardia cysts by amplification of heat shock-induced mRNA

Author:

Abbaszadegan M1,Huber M S1,Gerba C P1,Pepper I L1

Affiliation:

1. Quality Control & Research Laboratory, American Water Works Service Company, Inc., Belleville, Illinois 62220, USA.

Abstract

Primers obtained from gene sequences coding for heat shock proteins (HSP) were used to specifically detect enteric protozoans of the genus Giardia. The HSP primers amplified Giardia DNA or the corresponding RNA sequences obtained from lysed cysts and gave a 163-bp product. Since the presence of the product did not indicate whether the cysts were viable, these amplifications are a presence/absence test only. In contrast, amplification of heat shock-induced mRNA utilizing the same HSP primers was indicative of viable Giardia cysts. The limit of sensitivity of the presence/absence test was 1 cyst, whereas for the viability test it was 10 cysts. Thus, viable Giardia cysts can be rapidly and specifically detected with great sensitivity through the use of PCR amplifications.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Reference33 articles.

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2. Aggarwal A. P. Romans V. F. de la Cruz and T. E. Nash. 1988. Conserved sequences of the HSP gene family in Giardia lamblia p. 173-175. In P. M. Wallis and B. R. Hammond (ed.) Advances in Giardia research. University of Calgary Press Calgary Alberta Canada.

3. Drinking water transmission of giardiasis in the United States. Water Sci;Akin E. W.;Technol.,1986

4. Detection of viable Legionella pneumophila in water by polymerase chain reaction and gene probe methods;Bej A. K.;Appl. Environ. Microbiol.,1991

5. Giardia lamblia infections in Mongolian gerbils: an animal model;Belosevic M.;J. Infect. Dis.,1983

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