Enumeration of Legionella spp. from Environmental Sources

» ashleywcy - Enumeration of Legionella spp. from Environmental Sources

I'm an undergraduate currently involved in a project where I'm suppose to isolate and enumerate environmental strains of Legionella spp. by culture methods.

I'm having difficulty trying to lower the detection limits of the culture methods by concentrating the water samples. Furthermore, it is difficult to concentrate environmental water samples because there are a lot of sediments in the water which can clog up the filter.

Can anyone suggest other ways of lowering detection limits of the culture methods? Thank you very much.

-- posted by ashleywcy

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» NealC - Re: Enumeration of Legionella spp. from Environmental Sources

In response to message posted by ashleywcy:

I would lyophilize the concentrated water samples. You can dope some water with Legionella to see if they will survive the lyphophilization.

-- posted by NealC

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» NealC - Re: Enumeration of Legionella spp. from Environmental Sources

In response to message posted by ashleywcy:

Another way to go would be to eliminate the culturing entirely and use PCR to identify the few Legionella in the concentrated water samples.

-- posted by NealC

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» ashleywcy - PCR for detection

I would like to quantitate Legionella in the environmental samples. I had heard of RT-PCR, but I could not understand how it works.

-- posted by ashleywcy

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» RichardB91 - Concentrating Legionella

Hi there,

you can always give the sample a good mixing, and the use low speed centrifugation to get rid of sediment etc, the filter the supernatant. Alternatively just centrifuge and acid treat the sediment and then plate it. Either waLegionellala culture is very inaccurate anyway, so yoprobablyly wont lose much precision (since your probably working with at least +/- 30% already!)

cheers,

Richard

-- posted by RichardB91

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