Abstract
| Original language | English |
|---|---|
| Pages (from-to) | 7372-7377 |
| Number of pages | 5 |
| Journal | Applied and Environmental Microbiology |
| Volume | 70 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - Dec 2004 |
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Keywords
- quantification
- tetracycline resistance genes
- feedlot lagoons
- real-time PCR
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Quantification of tetracycline resistance genes in feedlot lagoons by real-time PCR. / Smith, M.S.; Yang, R.K.; Knapp, C.W.; Niu, Y.F.; Peak, N.; Hanfelt, M.M.; Galland, J.C.; Graham, D.W.
In: Applied and Environmental Microbiology, Vol. 70, No. 12, 12.2004, p. 7372-7377.Research output: Contribution to journal › Article
TY - JOUR
T1 - Quantification of tetracycline resistance genes in feedlot lagoons by real-time PCR
AU - Smith, M.S.
AU - Yang, R.K.
AU - Knapp, C.W.
AU - Niu, Y.F.
AU - Peak, N.
AU - Hanfelt, M.M.
AU - Galland, J.C.
AU - Graham, D.W.
PY - 2004/12
Y1 - 2004/12
N2 - A new real-time PCR method is presented that detects and quantifies three tetracycline resistance (Tc-r) genes [tet(O), tet(W), and tet(Q)] in mixed microbial communities resident in feedlot lagoon wastewater. Tc-r gene real-time TaqMan primer-probe sets were developed and optimized to quantify the Tc-r genes present in seven different cattle feedlot lagoons, to validate the method, and to assess whether resistance gene concentrations correlate with free-tetracycline levels in lagoon waters. The method proved to be sensitive across a wide range of gene concentrations and provided consistent and reproducible results from complex lagoon water samples. The log,0 of the sum of the three resistance gene concentrations was correlated with free-tetracycline levels (r(2) = 0.50, P<0.001; n = 18), with the geometric means of individual resistance concentrations ranging from 4- to 8.3-fold greater in lagoon samples with above-median tetracycline levels (>1.95 mug/liter by enzyme-linked immunosorbent assay techniques) than in below-median lagoon samples. Of the three Tc-r genes tested, tet(W) and tet(Q) were more commonly found in lagoon water samples. Successful development of this real-time PCR assay will permit other studies quantifying Tc-r gene numbers in environmental and other samples.
AB - A new real-time PCR method is presented that detects and quantifies three tetracycline resistance (Tc-r) genes [tet(O), tet(W), and tet(Q)] in mixed microbial communities resident in feedlot lagoon wastewater. Tc-r gene real-time TaqMan primer-probe sets were developed and optimized to quantify the Tc-r genes present in seven different cattle feedlot lagoons, to validate the method, and to assess whether resistance gene concentrations correlate with free-tetracycline levels in lagoon waters. The method proved to be sensitive across a wide range of gene concentrations and provided consistent and reproducible results from complex lagoon water samples. The log,0 of the sum of the three resistance gene concentrations was correlated with free-tetracycline levels (r(2) = 0.50, P<0.001; n = 18), with the geometric means of individual resistance concentrations ranging from 4- to 8.3-fold greater in lagoon samples with above-median tetracycline levels (>1.95 mug/liter by enzyme-linked immunosorbent assay techniques) than in below-median lagoon samples. Of the three Tc-r genes tested, tet(W) and tet(Q) were more commonly found in lagoon water samples. Successful development of this real-time PCR assay will permit other studies quantifying Tc-r gene numbers in environmental and other samples.
KW - quantification
KW - tetracycline resistance genes
KW - feedlot lagoons
KW - real-time PCR
UR - http://aem.asm.org/cgi/content/abstract/70/12/7372
UR - http://dx.doi.org/10.1128/AEM.70.12.7372-7377.2004
U2 - 10.1128/AEM.70.12.7372-7377.2004
DO - 10.1128/AEM.70.12.7372-7377.2004
M3 - Article
VL - 70
SP - 7372
EP - 7377
JO - Applied and Environmental Microbiology
JF - Applied and Environmental Microbiology
SN - 0099-2240
IS - 12
ER -