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Legionella
Sampling
Culture v PCR
       - By Nemco Utilities
(www.nemco-utilities.co.uk)
Nemco Utilities Limited


Legionella Sampling
Culture v PCR
The gold standard for Legionella testing has always
been the culture method (often referred to as the
standard reference method) whereby the bacteria are
filtered from a water sample, grown and then the
number of colony forming units (cfu) estimated.
However, this is a labour intensive process with a 12 day
turnaround. Given the penalties for non-compliance
and costs associated with taking water services out of
use for 12 days, the culture method is rapidly becoming
outdated.

    For more information about Legionella sampling or managing the risks associated with
                  Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
Nemco Utilities Limited


Legionella Sampling
Culture v PCR
A number of recent scientific advances have
revolutionised the face of Legionella analysis and now,
with a decent courier, samples can be turned around in
24 hours (4 in an emergency). Progress has been made
possible due to the use of Polymerase Chain Reaction
(PCR) technology. Rather than actually growing the
Legionella bacteria, PCR works by isolating fragments of
DNA from the water sample and amplifying them to a
detectable level. This process is a lot quicker, more
efficient and can largely be automated.

    For more information about Legionella sampling or managing the risks associated with
                  Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
Nemco Utilities Limited


Legionella Sampling
Culture v PCR
PCR has been of inestimable value especially for trend
analysis and when investigating outbreaks. It is not
without its problems. The initial stumbling block was that
of detecting ‘dead’ or ‘non viable’ cells, that would not
show up on culture. This has since been largely
overcome, but PCR in general terms still returns far more
positive results than the culture method. This leads on to
the next problem of reporting units and their correlation
with action levels.


     For more information about Legionella sampling or managing the risks associated with
                   Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
Nemco Utilities Limited


Legionella Sampling
Culture v PCR

The current Legionella guidance ACOP L8 contains
recommendations for ‘Action in the event of a positive
sample’ as below (for domestic water systems):
Legionella bacteria (cfu/                  ACTION REQUIRED (L8)
litre)
More than 100 but less than                Details >>
1000
More Than 1000                             Details >>

    For more information about Legionella sampling or managing the risks associated with
                  Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
Nemco Utilities Limited


Legionella Sampling
Culture v PCR
Legionella bacteria (cfu/ litre)             ACTION REQUIRED (L8)
More than 100 but less than                  Either:
1000                                         (a) If only one or two samples
                                             are positive, system should be
                                             re-sampled. If a similar count is
                                             found again, a review of the
                                             control measures and risk
                                             assessment should be carried
                                             out to identify any remedial
                                             actions.



     For more information about Legionella sampling or managing the risks associated with
                   Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
Nemco Utilities Limited


Legionella Sampling
Culture v PCR
Legionella bacteria (cfu/ litre)             ACTION REQUIRED (L8)
More than 100 but less than                  (b) If the majority of samples
1000                                         are positive, the system may
                                             be colonised, albeit at a low
                                             level,     with      legionella.
                                             Disinfection of the system
                                             should be considered but an
                                             immediate review of control
                                             measures and risk assessment
                                             should be carried out to
                                             identify any other remedial
                                             action required.
     For more information about Legionella sampling or managing the risks associated with
                   Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
Nemco Utilities Limited


Legionella Sampling
Culture v PCR
Legionella bacteria (cfu/ litre)             ACTION REQUIRED (L8)
More than 1000                               The system should be re-
                                             sampled and an immediate
                                             review of the control measures
                                             and risk assessment carried
                                             out to identify any remedial
                                             actions, including possible
                                             disinfection of the system.




     For more information about Legionella sampling or managing the risks associated with
                   Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
Nemco Utilities Limited


Legionella Sampling
Culture v PCR

The reporting units in this table refer entirely to colony
forming units (cfu), whereas PCR sample results are
returned in genomic units (GU). Therefore, if we get a
negative culture test but a positive PCR test – what
action to take? Or similarly, if PCR is used in isolation at
what point do you start spending large amounts of
money disinfecting water systems and taking services
out of action? There is no way of correlating the two
units and no set levels for genomic units.

     For more information about Legionella sampling or managing the risks associated with
                   Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
Nemco Utilities Limited


Legionella Sampling
Culture v PCR

Currently a large number of companies offering PCR,
back up the results with a culture test (at extra cost). It is
certainly in no doubt that future analysis will move
towards PCR (molecular photocopying as it is
sometimes known). In the meantime, we will be looking
to the HSE for guidance on the newly UKAS accredited
PCR testing method.



     For more information about Legionella sampling or managing the risks associated with
                   Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
Nemco Utilities Limited




Contact Details
Address                                      Email:
Nemco Utilities Ltd                           info@nemco-
12 Kempson Way                                 utilities.co.uk
Hillside Business Park                        sales@nemco-
Bury St Edmunds                                utilities.co.uk
Suffolk                                       accounts@nemco-
IP32 7EA                                       utilities.co.uk
                                             Website:
Telephone: 01284 724503                      http://www.nemco-
Fax No.: 01284 724826                        utilities.co.uk/
     For more information about Legionella sampling or managing the risks associated with
                   Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk

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Legionella sampling

  • 1. Legionella Sampling Culture v PCR - By Nemco Utilities (www.nemco-utilities.co.uk)
  • 2. Nemco Utilities Limited Legionella Sampling Culture v PCR The gold standard for Legionella testing has always been the culture method (often referred to as the standard reference method) whereby the bacteria are filtered from a water sample, grown and then the number of colony forming units (cfu) estimated. However, this is a labour intensive process with a 12 day turnaround. Given the penalties for non-compliance and costs associated with taking water services out of use for 12 days, the culture method is rapidly becoming outdated. For more information about Legionella sampling or managing the risks associated with Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
  • 3. Nemco Utilities Limited Legionella Sampling Culture v PCR A number of recent scientific advances have revolutionised the face of Legionella analysis and now, with a decent courier, samples can be turned around in 24 hours (4 in an emergency). Progress has been made possible due to the use of Polymerase Chain Reaction (PCR) technology. Rather than actually growing the Legionella bacteria, PCR works by isolating fragments of DNA from the water sample and amplifying them to a detectable level. This process is a lot quicker, more efficient and can largely be automated. For more information about Legionella sampling or managing the risks associated with Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
  • 4. Nemco Utilities Limited Legionella Sampling Culture v PCR PCR has been of inestimable value especially for trend analysis and when investigating outbreaks. It is not without its problems. The initial stumbling block was that of detecting ‘dead’ or ‘non viable’ cells, that would not show up on culture. This has since been largely overcome, but PCR in general terms still returns far more positive results than the culture method. This leads on to the next problem of reporting units and their correlation with action levels. For more information about Legionella sampling or managing the risks associated with Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
  • 5. Nemco Utilities Limited Legionella Sampling Culture v PCR The current Legionella guidance ACOP L8 contains recommendations for ‘Action in the event of a positive sample’ as below (for domestic water systems): Legionella bacteria (cfu/ ACTION REQUIRED (L8) litre) More than 100 but less than Details >> 1000 More Than 1000 Details >> For more information about Legionella sampling or managing the risks associated with Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
  • 6. Nemco Utilities Limited Legionella Sampling Culture v PCR Legionella bacteria (cfu/ litre) ACTION REQUIRED (L8) More than 100 but less than Either: 1000 (a) If only one or two samples are positive, system should be re-sampled. If a similar count is found again, a review of the control measures and risk assessment should be carried out to identify any remedial actions. For more information about Legionella sampling or managing the risks associated with Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
  • 7. Nemco Utilities Limited Legionella Sampling Culture v PCR Legionella bacteria (cfu/ litre) ACTION REQUIRED (L8) More than 100 but less than (b) If the majority of samples 1000 are positive, the system may be colonised, albeit at a low level, with legionella. Disinfection of the system should be considered but an immediate review of control measures and risk assessment should be carried out to identify any other remedial action required. For more information about Legionella sampling or managing the risks associated with Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
  • 8. Nemco Utilities Limited Legionella Sampling Culture v PCR Legionella bacteria (cfu/ litre) ACTION REQUIRED (L8) More than 1000 The system should be re- sampled and an immediate review of the control measures and risk assessment carried out to identify any remedial actions, including possible disinfection of the system. For more information about Legionella sampling or managing the risks associated with Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
  • 9. Nemco Utilities Limited Legionella Sampling Culture v PCR The reporting units in this table refer entirely to colony forming units (cfu), whereas PCR sample results are returned in genomic units (GU). Therefore, if we get a negative culture test but a positive PCR test – what action to take? Or similarly, if PCR is used in isolation at what point do you start spending large amounts of money disinfecting water systems and taking services out of action? There is no way of correlating the two units and no set levels for genomic units. For more information about Legionella sampling or managing the risks associated with Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
  • 10. Nemco Utilities Limited Legionella Sampling Culture v PCR Currently a large number of companies offering PCR, back up the results with a culture test (at extra cost). It is certainly in no doubt that future analysis will move towards PCR (molecular photocopying as it is sometimes known). In the meantime, we will be looking to the HSE for guidance on the newly UKAS accredited PCR testing method. For more information about Legionella sampling or managing the risks associated with Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk
  • 11. Nemco Utilities Limited Contact Details Address Email: Nemco Utilities Ltd  info@nemco- 12 Kempson Way utilities.co.uk Hillside Business Park  sales@nemco- Bury St Edmunds utilities.co.uk Suffolk  accounts@nemco- IP32 7EA utilities.co.uk Website: Telephone: 01284 724503 http://www.nemco- Fax No.: 01284 724826 utilities.co.uk/ For more information about Legionella sampling or managing the risks associated with Legionella, please contact Nemco Utilities at www.nemco-utilities.co.uk