Comments on the routine testing of water samples for Legionella sp.

Taylor, Rebecca L.; Ndabula, Christopher; Robinson, J. S.

doi:10.1016/0043-1354(94)00308-t

Cited by 2 publications

(2 citation statements)

References 5 publications

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“…On the basis of these results, it can be concluded that the heat pretreatment method is preferable to that using acid as, under the various conditions of contamination, it gave results that were better, or at least equivalent. This conclusion is in agreement with those of other studies Verran et al 1995;De Luca et al 1999). The direct inoculum of untreated samples showed low sensitivity and low speci®city, but it nevertheless provided values nearer to the real numbers of Legionellae present in the samples.…”

Section: Discussionsupporting

confidence: 94%

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Comparison of selective procedures for isolation and enumeration ofLegionellaspecies from hot water systems

Leoni

Legnani

2001

Journal of Applied Microbiology

112

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E. LEONI AND P.P. LEGNANI. 2001. Various sample pre‐treatment techniques and different growth media for the isolation of Legionellae from hot water supplies in public buildings were compared. A total of 102 hot water samples from taps and showers was examined. The highest recovery frequency was obtained with the heat pre‐treatment method and using the selective medium GVPC. However, the results differed according to the concentration of legionellas. In the case of low plate counts (≤5000 cfu l−1), the heat pre‐treatment technique gave a significantly higher percentage of positive samples compared with other techniques (P < 0·05). With increasing concentration, the differences between the procedures decreased until they became statistically not significant for concentrations above 50 000 cfu l−1. The direct inoculum method allowed a significantly higher detection of concentrations (P < 0·001) compared with heat and acid decontamination methods, which brought about a 67–68% reduction in detectable Legionellae. Heat decontamination techniques show greater sensitivity and specificity. However, they underestimate the number of legionellas. In environmental surveillance programmes, this underestimate must be taken into consideration when assessing the health risk.

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Section: Discussionsupporting

confidence: 94%

“…This conclusion is in agreement with those of other studies (Groothuis et al . 1985; Verran et al . 1995; De Luca et al .…”

Section: Discussionmentioning

confidence: 99%

Comparison of selective procedures for isolation and enumeration ofLegionellaspecies from hot water systems

Leoni

Legnani

2001

Journal of Applied Microbiology

112

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Enhancement of Legionella pneumophila Culture Isolation from Microenvironments by Macrophage Infectivity Potentiator (mip) Gene‐Specific Nested Polymerase Chain Reaction

Nintasen

Utrarachkij

Siripanichgon

et al. 2007

Microbiology and Immunology

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The combination of a Legionella pneumophila culture isolation technique and macrophage infectivity potentiator (mip) gene‐specific nested polymerase chain reaction (PCR) is pivotal for effective routine use in an environmental water system laboratory. Detection of Legionella organisms in 169 environmental samples was performed by using modified buffered charcoal yeast extract (MBCYE) agar for conventional culture. Nested PCR specific for L. pneumophila was performed using boiled genomic DNA extracts from filtered and Chelex 100‐treated water samples, or by using silica‐gel membrane spin column‐eluted DNA from concentrated pond, canal and river samples. Overall, the nested PCR was twelvefold more sensitive than the culture method. The target amplicons (471 basepairs) of all 4 biochemically characterized L. pneumophila isolates were sequenced. They had homology at the DNA and protein levels to 3′ proximity of the mip‐coding gene of L. pneumophila deposited in genome databases. EcoRI‐ or KpnI‐digested PCR fragments with expected sizes were also confirmed in all 52 PCR‐positive samples that were isolated from cooling towers and condenser drains. Viable but nonculturable L. pneumophila might have been present in 48 PCR‐positive samples. This study demonstrates that detection of the genetically stable mip gene by nested PCR with a modified process of water sample preparation can be rapidly and effectively used to enhance isolation of the L. pneumophila taxon from microenvironments.

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Comments on the routine testing of water samples for Legionella sp.

Cited by 2 publications

References 5 publications

Comparison of selective procedures for isolation and enumeration ofLegionellaspecies from hot water systems

Comparison of selective procedures for isolation and enumeration ofLegionellaspecies from hot water systems

Enhancement of Legionella pneumophila Culture Isolation from Microenvironments by Macrophage Infectivity Potentiator (mip) Gene‐Specific Nested Polymerase Chain Reaction

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