This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Mahbubani, M H Articles by Atlas, R M Search for Related Content PubMed PubMed Citation Articles by Mahbubani, M H Articles by Atlas, R M Agricola Articles by Mahbubani, M H Articles by Atlas, R M

 Previous Article  |  Next Article 

Appl Environ Microbiol. 1991 December; 57(12): 3456-3461

Detection of Giardia cysts by using the polymerase chain reaction and distinguishing live from dead cysts.

Department of Biology, University of Louisville, Kentucky 40292.

ABSTRACT

A method was developed for the detection of Giardia cysts by using the polymerase chain reaction (PCR) and the giardin gene as the target. DNA amplification by PCR, using giardin DNA as the target, resulted in detection of both live and dead cysts. When giardin mRNA was used as the target, the ability to amplify cDNA by PCR depended on the mode of killing. Cysts killed by freezing were not detected by PCR when giardin mRNA was the target. Cysts killed by heating or exposure to monochloramine, however, gave positive detection signals for both DNA and giardin mRNA targets. The amount of giardin mRNA and total RNA was significantly increased in live cysts following the induction of excystation. Cysts killed by freezing, heating, or exposure to monochloramine did not show a change in RNA content. The detection of the giardin gene by PCR permits a sensitive and specific diagnosis for Giardia spp. Discrimination between live and dead cysts can be made by measuring the amounts of RNA or PCR-amplified product from the giardin mRNA target before and after the induction of excystation.

This article has been cited by other articles:

• Guy, R. A., Payment, P., Krull, U. J., Horgen, P. A. (2003). Real-Time PCR for Quantification of Giardia and Cryptosporidium in Environmental Water Samples and Sewage. Appl. Environ. Microbiol. 69: 5178-5185 [Abstract] [Full Text]  
• Kaucner, C., Stinear, T. (1998). Sensitive and Rapid Detection of Viable Giardia Cysts and Cryptosporidium parvum Oocysts in Large-Volume Water Samples with Wound Fiberglass Cartridge Filters and Reverse Transcription-PCR. Appl. Environ. Microbiol. 64: 1743-1749 [Abstract] [Full Text]