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Chapter 106 : Detection from Water Samples by Real-Time PCR

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Detection from Water Samples by Real-Time PCR, Page 1 of 2

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Abstract:

This chapter deals about the direct detection of from hospital water samples by commercial real-time (RT)-PCR system, that offers rapid results and reduced risk of cross-contamination. The preliminary results of 61 water samples from 6 hospitals and 15 water samples from air conditioning systems are presented and compared with those of the isolation method. Water samples were concentrated by filtration through cellulose acetate membrane filter and resuspended in 10 ml of the same water. The principle steps were cell lysis with proteinase K, purification on spin column and elution. Culture was positive in 28 (45.9%) hospital water samples, and the strains were identified as serogroup 1, 3, 6, and 2-14 and spp. The results of qualitative RT-PCR were as follows: spp. was positive (detection limit: 133 GU/liter) in 60 (98.4%) samples and was positive in 44 (72.1%). The results of culture and qualitative analysis for by RT-PCR are shown. All hospitals examined showed some positive samples for culture and PCR, except for one that was culture negative and PCR positive. All water samples from air conditioning systems were culture negative, but RT-PCR was positive in 15 water samples for spp. and in 2 for . In conclusion, detection by RT-PCR from environmental methods seems to be promising. However, at the moment, correlation between the method-result interpretations and public health significance need further evaluations.

Citation: Franzin L, Cabodi D, Bonfrate N. 2006. Detection from Water Samples by Real-Time PCR, p 446-448. 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.ch106

Key Concept Ranking

Legionella pneumophila
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Real-Time PCR
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FIGURE 1

Comparison of culture and quantitative PCR methods for enumeration of .

Citation: Franzin L, Cabodi D, Bonfrate N. 2006. Detection from Water Samples by Real-Time PCR, p 446-448. 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.ch106
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References

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1. Ballard, A. L.,, N. K. Fry,, L. Chan,, S. B. Sur-man,, J. V. Lee,, T. G. Harrison, and, K. J. Towner. 2000. Detection of Legionella pneumophila using a real-time PCR hybridization assay. J. Clin. Microbiol. 38:42154218.
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Tables

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TABLE 1

Results of culture and qualitative analysis for by RT-PCR

Citation: Franzin L, Cabodi D, Bonfrate N. 2006. Detection from Water Samples by Real-Time PCR, p 446-448. 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.ch106

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