PMID: 9425249Mar 14, 1998Paper

Detection of Giardia in environmental waters by immuno-PCR amplification methods

Current Microbiology
M H MahbubaniA K Bej

Abstract

Genomic DNA was extracted either directly from Giardia muris cysts seeded into environmental surface waters or from cysts isolated by immunomagnetic beads (IMB). A 0.171-kbp segment of the giardin gene was PCR-amplified following "direct extraction" of Giardia DNA from seeded Cahaba river water concentrate with moderate turbidity (780 JTU's), but DNA purified from seeded Colorado river water concentrates with high turbidity (2 x 10(5) JTUs) failed to amplify. However, if the cysts were first separated by the IMB approach from seeded Cahaba or Colorado river waters, and the DNA released by a freeze-boil Chelex(R)100 treatment, detection of G. muris by PCR amplification could be achieved at a sensitivity of 3 x 10(0) or 3 x 10(1) cysts/ml, respectively. If, however, the G. muris cysts used to seed even moderately turbid river waters (780 JTUs) were formalin treated (which is conventionally used for microscopic examination), neither direct extraction nor IMB purification methods yielded amplifiable DNA. Use of immunomagnetic beads to separate Giardia cysts from complex matrices of environmental surface waters followed by DNA release and PCR amplification of the target giardin gene improved the reliability of detection of this path...Continue Reading

Citations

May 15, 2012·Analytical and Bioanalytical Chemistry·Ivo SafarikMirka Safarikova
Aug 9, 2005·International Journal of Food Microbiology·David Dawson
Mar 9, 2004·Journal of Microbiological Methods·Isabelle BertrandJanine Schwartzbrod
Sep 6, 2003·Applied and Environmental Microbiology·Rebecca A GuyPaul A Horgen
Jan 9, 2001·Clinical Microbiology Reviews·T B Gardner, D R Hill
Oct 15, 2005·Clinical Microbiology Reviews·Daniel V LimMarianne F Kramer
Mar 11, 1999·Experimental Parasitology·M F HeyworthT W Sell
Jan 5, 2002·Expert Opinion on Investigational Drugs·T G Mank, J O Zaat
Aug 12, 2009·Environmental Research·Yvonne A L LimHuw V Smith
Mar 28, 2008·Memórias do Instituto Oswaldo Cruz·Marta C MinvielleJuan A Basualdo
May 11, 2002·Critical Reviews in Microbiology·J Theron, T E Cloete
Dec 7, 2017·Korean journal of medical education·Sanghee Yeo, Bong Hyun Chang
Mar 4, 2000·The Journal of Parasitology·L McIntyreJ L Isaac-Renton
Sep 4, 2021·Journal of Microbiological Methods·Pengbo LiuChristine L Moe
Jun 29, 1999·Analytical Chemistry·S D Richardson

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