NTM qPCR

NTM qPCR

NTM qPCR (Presence / Absence)

Significance

This rapid molecular screening method for detection of nontuberculous mycobacteria (NTM) test utilizes quantitative polymerase chain reaction (qPCR) to amplify and detect low level amounts of DNA specific to NTM, thereby reducing the standard turnaround time of 6-8 weeks for culture down to 2-4 days for qPCR.  Results are provided as presence/absence with an approximate range for low, medium and high levels of detection. When time is a factor, this test provides fast results to enable a faster response!  This means less equipment downtime, less water restrictions, and a healthier system.

When species identification is important, we recommend pairing qPCR with culture as qPCR will not provide information on individual species present.  Culture not only enables live organism speciation but also isolate retention for later comparisons such as those required in clinical case investigations.

The procedure used for qPCR follows International Standard Organization (ISO) standard 12869-2019 with minor modifications. This standard qPCR cannot discriminate the genome of live vs dead/impaired organisms and may also detect NTM that are not culturable. However, this detection of all DNA present contributes to this test’s high predictive power as a negative screen because a negative result indicates that no NTM (live or dead) is present.

.Test Method

• Water samples are concentrated by filtration.
• DNA is extracted using the Qiagen DNeasy Power Water Kit.
• Quantitative polymerase chain reaction (qPCR) for detection of non-tuberculous mycobacteria.
• Qualitative (+/-) result with 3 approximate detection levels.

Turn Around Time
2–4 days

Test Code
145

Sample Transport
See Sampling and Shipping page for instructions.

References

qPCR Application Guide – Experimental Overview, Protocol, Troubleshooting. 3rd Edition. 2011-2012. Integrated DNA Technologies.

Qiagen DNeasy PowerWater DNA Isolation Kit Instruction Manual.

Applied Biosystems. Quantstudio 6 and 7 Flex Real-Time PCR System Software. 2013. Booklets 2 and 5.

ISO Standard 12869: 2019. Water Quality – Detection and quantification of Legionella spp. by concentration and genic amplification by quantitative polymerase chain reaction (qPCR)

Beumer A, King D, Donohue M, Mistry J, Covert T, Pfaller S. Detection of mycobacterium avium subsp. paratuberculosis in drinking water and biofilms by quantitative PCR. Applied and Environmental Microbiology. 2010 Nov; 76 (21):7367–7370. doi:10.1128/AEM.00730-10

Accreditations and Proficiency
See Accreditations and Proficiency page for information on our accreditations and Legionella testing proficiency programs.