Evaluation of a Method to Detect <i>Giardia</i> and <i>Cryptosporidium</i> in Water

LeChevallier, MW; Norton, WD; Rg, Lee

doi:10.1520/stp17618s

Monitoring Water in the 1990's: Meeting New Challenges 1991

DOI: 10.1520/stp17618s

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Evaluation of a Method to Detect Giardia and Cryptosporidium in Water

MW LeChevallier

WD Norton

Lee Rg

Abstract: The current study outlines a technique to simultaneously detect Giardia and Cryptosporidium in raw and tap water samples. The influence of various procedural steps, including filtration, elution, flotation, filter mounting and microscopy, was determined for the method. Overall, the method recovered 48% to 69% of the Giardia and 25% to 42% of the Cryptosporidium in spiked water samples. The method had an averaged detection limit of 1 organism/100 L of tap water. Use of phase-contrast and Nomarski differential i… Show more

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Cited by 13 publications

References 13 publications

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Examining Relationships Between Particle Counts and Giardia, Cryptosporidium, and Turbidity

LeChevallier¹,

Norton²

1992

Journal AWWA

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This study examines concentrations of Giardia and Cryptosporidium, turbidity, and particles in raw and filtered waters as indicators of treatment plant efficiency. Results indicate that the probability of detecting cysts and oocysts in finished water supplies depends on the number of organisms in the raw water. Particle counts are useful indicators of plant performance and can indicate when filters are not functioning properly.

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Examining Relationships Between Particle Counts and Giardia, Cryptosporidium, and Turbidity

LeChevallier¹,

Norton²

1992

Journal AWWA

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Using flow cytometry to detect protozoa

Hoffman¹,

Standridge²,

Prieve³

et al. 1997

Journal AWWA

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For routine detection of Cryptosporidium and Giardia, flow cytometry with cell sorting bests IFA in terms of relative sensitivity as well as average cost, assay volume, assay time, and turnaround time. Flow cytometric cell sorting (FCCS), followed by microscopic verification, was evaluated as an alternative to immunofluorescence assay (IFA). Two hundred and sixty‐two water samples were stained with either indirectly conjugated or directly conjugated anti‐Cryptosporidium and anti‐Giardia antibodies and were examined by IFA and FCCS, respectively. Relative sensitivities of each assay were calculated by dividing the number of positive samples detected by one assay by the number of positive samples detected by either assay. When samples analyzed within five months of the initial IFA analysis were compared, the relative sensitivities of FCCS for detection of Cryptosporidium and Giardia were 94.1 and 78.6 percent, respectively, whereas the relative sensitivities using the IFA were 35.3 and 61.4 percent. Identification of Cryptosporidium and Giardia in water samples using FCCS takes less time, costs less, and analyzes a greater sample volume than the IFA. Given the nearly threefold increase in relative sensitivity for Cryptosporidium detection and the equal relative sensitivities for Giardia detection, the authors support the use of FCCS for routine detection of Cryptosporidium and Giardia in environmental samples.

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Detection of Giardia in Environmental Waters by Immuno-PCR Amplification Methods

Mahbubani

Schaefer

Jones

et al. 1998

Current Microbiology

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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 pathogen with the required sensitivity.

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Evaluation of a Method to Detect Giardia and Cryptosporidium in Water

Cited by 13 publications

References 13 publications

Examining Relationships Between Particle Counts and Giardia, Cryptosporidium, and Turbidity

Examining Relationships Between Particle Counts and Giardia, Cryptosporidium, and Turbidity

Using flow cytometry to detect protozoa

Detection of Giardia in Environmental Waters by Immuno-PCR Amplification Methods

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