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Chapter 108 : A Novel and Rapid Detection System for Water Analysis

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A Novel and Rapid Detection System for Water Analysis, Page 1 of 2

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Abstract:

The usefulness of sampling waters for the detection of remains a point of discussion among researchers and industry for a variety of reasons. This chapter describes a novel detection system utilizing two liquid media (medium 1 and medium 2, formulations are currently under provisional patent applications) and real-time PCR that is capable of detecting viable in a variety of waters within 2 to 3 days. Over 30 strains of have been tested by this method, and the real-time PCR detection system has been validated by authors' laboratory. Real-time PCR utilizing 16S rRNA primers was performed on a Corbett Rotor-Gene 3000 utilizing SYTO9 as a DNA intercalating dye for PCR. The GMS method was challenged with 82 field samples (cooling tower, evaporative tower, and potable waters) and run in parallel to AS/NZS 3896:1998. Comparison of methods using field samples showed that the GMS method is 100% specific (no false positives) and 92% sensitive, with a single discrepant sample. The GMS method described is rapid, simple, cost-effective, less laborious than conventional culture, and easy to implement in any modern day microbiology laboratory.

Citation: Giglio S, T. Monis P, P. Saint C. 2006. A Novel and Rapid Detection System for Water Analysis, p 453-455. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch108

Key Concept Ranking

Legionella pneumophila
0.76785713
Real-Time PCR
0.6517858
0.76785713
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Figures

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FIGURE 1

Flow chart of the new GMS method.

Citation: Giglio S, T. Monis P, P. Saint C. 2006. A Novel and Rapid Detection System for Water Analysis, p 453-455. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch108
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References

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1. Anonymous. 1998. Waters: Examination for Legionellae AS/NZS 3896:1998. Standards Australia, North Sydney, NSW, Australia.
2. Arghros, M.,, G. Douglass,, M. Locher,, P. Mugg,, D. C. Myatt,, T. Olma,, A. Scholtes, and, I. Wilkinson. 2004. Guidelines for Assuring Quality of Food and Water Microbiological Culture Media. Australian Society for Microbiology, Media Quality Control Special Interest Group.
3. Broadbent, C. 1996. Guidance for the Control of Legionella. National Environmental Health Forum Monographs, Water Series no. 1.
4. Giglio,, S.,, P. T. Monis, and, C. P. Saint. 2005. Legionella confirmation using real-time PCR and SYTO9 is an alternative to current methodology. Appl. Environ. Microbiol. 71:89448948.
5. Monis, P. T.,, S. Giglio, and, C. P. Saint. 2005. Comparison of SYTO9 and SYBR Green I for real-time polymerase chain reaction and investigation of the effect of dye concentration on amplification and DNA melting curve analysis. Anal. Biochem. 340:2434.
6. Torii, K.,, Y. Iinuma,, M. Ichikawa,, K. Kato,, M. Koide,, H. Baba,, R. Suzuki, and, M. Ohta. 2003. A case of nosocomial Legionella pneumophila pneumonia. Jpn. J. Infect. Dis. 56:101102.

Tables

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TABLE 1

Validation of the GMS method against AS/NZS 4659 and AS/NZ 3896:1998

Citation: Giglio S, T. Monis P, P. Saint C. 2006. A Novel and Rapid Detection System for Water Analysis, p 453-455. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch108

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