Evaluation of the repeatability and reproducibility of a suite of qPCR-based microbial source tracking methods
Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowl...
Ausführliche Beschreibung
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| Autor*in: |
Ebentier, Darcy L. [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2013transfer abstract |
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| Schlagwörter: |
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| Umfang: |
10 |
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| Übergeordnetes Werk: |
Enthalten in: Matches, mismatches and priorities of pathways from a climate-resilient development perspective in the mountains of Nepal - Pandey, Avash ELSEVIER, 2021, a journal of the International Association on Water Quality (IAWQ), Amsterdam [u.a.] |
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| Übergeordnetes Werk: |
volume:47 ; year:2013 ; number:18 ; day:15 ; month:11 ; pages:6839-6848 ; extent:10 |
| Links: |
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| DOI / URN: |
10.1016/j.watres.2013.01.060 |
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| Katalog-ID: |
ELV021577765 |
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| 245 | 1 | 0 | |a Evaluation of the repeatability and reproducibility of a suite of qPCR-based microbial source tracking methods |
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| 520 | |a Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. | ||
| 520 | |a Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. | ||
| 650 | 7 | |a Repeatability |2 Elsevier | |
| 650 | 7 | |a Fecal pollution |2 Elsevier | |
| 650 | 7 | |a Microbial source tracking |2 Elsevier | |
| 650 | 7 | |a Reproducibility |2 Elsevier | |
| 650 | 7 | |a qPCR |2 Elsevier | |
| 700 | 1 | |a Hanley, Kaitlyn T. |4 oth | |
| 700 | 1 | |a Cao, Yiping |4 oth | |
| 700 | 1 | |a Badgley, Brian D. |4 oth | |
| 700 | 1 | |a Boehm, Alexandria B. |4 oth | |
| 700 | 1 | |a Ervin, Jared S. |4 oth | |
| 700 | 1 | |a Goodwin, Kelly D. |4 oth | |
| 700 | 1 | |a Gourmelon, Michèle |4 oth | |
| 700 | 1 | |a Griffith, John F. |4 oth | |
| 700 | 1 | |a Holden, Patricia A. |4 oth | |
| 700 | 1 | |a Kelty, Catherine A. |4 oth | |
| 700 | 1 | |a Lozach, Solen |4 oth | |
| 700 | 1 | |a McGee, Charles |4 oth | |
| 700 | 1 | |a Peed, Lindsay A. |4 oth | |
| 700 | 1 | |a Raith, Meredith |4 oth | |
| 700 | 1 | |a Ryu, Hodon |4 oth | |
| 700 | 1 | |a Sadowsky, Michael J. |4 oth | |
| 700 | 1 | |a Scott, Elizabeth A. |4 oth | |
| 700 | 1 | |a Domingo, Jorge Santo |4 oth | |
| 700 | 1 | |a Schriewer, Alexander |4 oth | |
| 700 | 1 | |a Sinigalliano, Christopher D. |4 oth | |
| 700 | 1 | |a Shanks, Orin C. |4 oth | |
| 700 | 1 | |a Van De Werfhorst, Laurie C. |4 oth | |
| 700 | 1 | |a Wang, Dan |4 oth | |
| 700 | 1 | |a Wuertz, Stefan |4 oth | |
| 700 | 1 | |a Jay, Jennifer A. |4 oth | |
| 773 | 0 | 8 | |i Enthalten in |n Elsevier Science |a Pandey, Avash ELSEVIER |t Matches, mismatches and priorities of pathways from a climate-resilient development perspective in the mountains of Nepal |d 2021 |d a journal of the International Association on Water Quality (IAWQ) |g Amsterdam [u.a.] |w (DE-627)ELV006716016 |
| 773 | 1 | 8 | |g volume:47 |g year:2013 |g number:18 |g day:15 |g month:11 |g pages:6839-6848 |g extent:10 |
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2013transfer abstract |
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2013 |
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10.1016/j.watres.2013.01.060 doi GBVA2013001000018.pica (DE-627)ELV021577765 (ELSEVIER)S0043-1354(13)00548-4 DE-627 ger DE-627 rakwb eng 550 550 DE-600 333.7 320 VZ Ebentier, Darcy L. verfasserin aut Evaluation of the repeatability and reproducibility of a suite of qPCR-based microbial source tracking methods 2013transfer abstract 10 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. Repeatability Elsevier Fecal pollution Elsevier Microbial source tracking Elsevier Reproducibility Elsevier qPCR Elsevier Hanley, Kaitlyn T. oth Cao, Yiping oth Badgley, Brian D. oth Boehm, Alexandria B. oth Ervin, Jared S. oth Goodwin, Kelly D. oth Gourmelon, Michèle oth Griffith, John F. oth Holden, Patricia A. oth Kelty, Catherine A. oth Lozach, Solen oth McGee, Charles oth Peed, Lindsay A. oth Raith, Meredith oth Ryu, Hodon oth Sadowsky, Michael J. oth Scott, Elizabeth A. oth Domingo, Jorge Santo oth Schriewer, Alexander oth Sinigalliano, Christopher D. oth Shanks, Orin C. oth Van De Werfhorst, Laurie C. oth Wang, Dan oth Wuertz, Stefan oth Jay, Jennifer A. oth Enthalten in Elsevier Science Pandey, Avash ELSEVIER Matches, mismatches and priorities of pathways from a climate-resilient development perspective in the mountains of Nepal 2021 a journal of the International Association on Water Quality (IAWQ) Amsterdam [u.a.] (DE-627)ELV006716016 volume:47 year:2013 number:18 day:15 month:11 pages:6839-6848 extent:10 https://doi.org/10.1016/j.watres.2013.01.060 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U AR 47 2013 18 15 1115 6839-6848 10 045F 550 |
| spelling |
10.1016/j.watres.2013.01.060 doi GBVA2013001000018.pica (DE-627)ELV021577765 (ELSEVIER)S0043-1354(13)00548-4 DE-627 ger DE-627 rakwb eng 550 550 DE-600 333.7 320 VZ Ebentier, Darcy L. verfasserin aut Evaluation of the repeatability and reproducibility of a suite of qPCR-based microbial source tracking methods 2013transfer abstract 10 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. Repeatability Elsevier Fecal pollution Elsevier Microbial source tracking Elsevier Reproducibility Elsevier qPCR Elsevier Hanley, Kaitlyn T. oth Cao, Yiping oth Badgley, Brian D. oth Boehm, Alexandria B. oth Ervin, Jared S. oth Goodwin, Kelly D. oth Gourmelon, Michèle oth Griffith, John F. oth Holden, Patricia A. oth Kelty, Catherine A. oth Lozach, Solen oth McGee, Charles oth Peed, Lindsay A. oth Raith, Meredith oth Ryu, Hodon oth Sadowsky, Michael J. oth Scott, Elizabeth A. oth Domingo, Jorge Santo oth Schriewer, Alexander oth Sinigalliano, Christopher D. oth Shanks, Orin C. oth Van De Werfhorst, Laurie C. oth Wang, Dan oth Wuertz, Stefan oth Jay, Jennifer A. oth Enthalten in Elsevier Science Pandey, Avash ELSEVIER Matches, mismatches and priorities of pathways from a climate-resilient development perspective in the mountains of Nepal 2021 a journal of the International Association on Water Quality (IAWQ) Amsterdam [u.a.] (DE-627)ELV006716016 volume:47 year:2013 number:18 day:15 month:11 pages:6839-6848 extent:10 https://doi.org/10.1016/j.watres.2013.01.060 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U AR 47 2013 18 15 1115 6839-6848 10 045F 550 |
| allfields_unstemmed |
10.1016/j.watres.2013.01.060 doi GBVA2013001000018.pica (DE-627)ELV021577765 (ELSEVIER)S0043-1354(13)00548-4 DE-627 ger DE-627 rakwb eng 550 550 DE-600 333.7 320 VZ Ebentier, Darcy L. verfasserin aut Evaluation of the repeatability and reproducibility of a suite of qPCR-based microbial source tracking methods 2013transfer abstract 10 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. Repeatability Elsevier Fecal pollution Elsevier Microbial source tracking Elsevier Reproducibility Elsevier qPCR Elsevier Hanley, Kaitlyn T. oth Cao, Yiping oth Badgley, Brian D. oth Boehm, Alexandria B. oth Ervin, Jared S. oth Goodwin, Kelly D. oth Gourmelon, Michèle oth Griffith, John F. oth Holden, Patricia A. oth Kelty, Catherine A. oth Lozach, Solen oth McGee, Charles oth Peed, Lindsay A. oth Raith, Meredith oth Ryu, Hodon oth Sadowsky, Michael J. oth Scott, Elizabeth A. oth Domingo, Jorge Santo oth Schriewer, Alexander oth Sinigalliano, Christopher D. oth Shanks, Orin C. oth Van De Werfhorst, Laurie C. oth Wang, Dan oth Wuertz, Stefan oth Jay, Jennifer A. oth Enthalten in Elsevier Science Pandey, Avash ELSEVIER Matches, mismatches and priorities of pathways from a climate-resilient development perspective in the mountains of Nepal 2021 a journal of the International Association on Water Quality (IAWQ) Amsterdam [u.a.] (DE-627)ELV006716016 volume:47 year:2013 number:18 day:15 month:11 pages:6839-6848 extent:10 https://doi.org/10.1016/j.watres.2013.01.060 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U AR 47 2013 18 15 1115 6839-6848 10 045F 550 |
| allfieldsGer |
10.1016/j.watres.2013.01.060 doi GBVA2013001000018.pica (DE-627)ELV021577765 (ELSEVIER)S0043-1354(13)00548-4 DE-627 ger DE-627 rakwb eng 550 550 DE-600 333.7 320 VZ Ebentier, Darcy L. verfasserin aut Evaluation of the repeatability and reproducibility of a suite of qPCR-based microbial source tracking methods 2013transfer abstract 10 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. Repeatability Elsevier Fecal pollution Elsevier Microbial source tracking Elsevier Reproducibility Elsevier qPCR Elsevier Hanley, Kaitlyn T. oth Cao, Yiping oth Badgley, Brian D. oth Boehm, Alexandria B. oth Ervin, Jared S. oth Goodwin, Kelly D. oth Gourmelon, Michèle oth Griffith, John F. oth Holden, Patricia A. oth Kelty, Catherine A. oth Lozach, Solen oth McGee, Charles oth Peed, Lindsay A. oth Raith, Meredith oth Ryu, Hodon oth Sadowsky, Michael J. oth Scott, Elizabeth A. oth Domingo, Jorge Santo oth Schriewer, Alexander oth Sinigalliano, Christopher D. oth Shanks, Orin C. oth Van De Werfhorst, Laurie C. oth Wang, Dan oth Wuertz, Stefan oth Jay, Jennifer A. oth Enthalten in Elsevier Science Pandey, Avash ELSEVIER Matches, mismatches and priorities of pathways from a climate-resilient development perspective in the mountains of Nepal 2021 a journal of the International Association on Water Quality (IAWQ) Amsterdam [u.a.] (DE-627)ELV006716016 volume:47 year:2013 number:18 day:15 month:11 pages:6839-6848 extent:10 https://doi.org/10.1016/j.watres.2013.01.060 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U AR 47 2013 18 15 1115 6839-6848 10 045F 550 |
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In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. Repeatability Elsevier Fecal pollution Elsevier Microbial source tracking Elsevier Reproducibility Elsevier qPCR Elsevier Hanley, Kaitlyn T. oth Cao, Yiping oth Badgley, Brian D. oth Boehm, Alexandria B. oth Ervin, Jared S. oth Goodwin, Kelly D. oth Gourmelon, Michèle oth Griffith, John F. oth Holden, Patricia A. oth Kelty, Catherine A. oth Lozach, Solen oth McGee, Charles oth Peed, Lindsay A. oth Raith, Meredith oth Ryu, Hodon oth Sadowsky, Michael J. oth Scott, Elizabeth A. oth Domingo, Jorge Santo oth Schriewer, Alexander oth Sinigalliano, Christopher D. oth Shanks, Orin C. oth Van De Werfhorst, Laurie C. oth Wang, Dan oth Wuertz, Stefan oth Jay, Jennifer A. oth Enthalten in Elsevier Science Pandey, Avash ELSEVIER Matches, mismatches and priorities of pathways from a climate-resilient development perspective in the mountains of Nepal 2021 a journal of the International Association on Water Quality (IAWQ) Amsterdam [u.a.] (DE-627)ELV006716016 volume:47 year:2013 number:18 day:15 month:11 pages:6839-6848 extent:10 https://doi.org/10.1016/j.watres.2013.01.060 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U AR 47 2013 18 15 1115 6839-6848 10 045F 550 |
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evaluation of the repeatability and reproducibility of a suite of qpcr-based microbial source tracking methods |
| title_auth |
Evaluation of the repeatability and reproducibility of a suite of qPCR-based microbial source tracking methods |
| abstract |
Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. |
| abstractGer |
Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. |
| abstract_unstemmed |
Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thoroughly evaluated. Knowledge of factors influencing PCR in different laboratories is vital to future technology transfer for use of MST methods as a tool for water quality management. In this study, a blinded set of 64 filters (containing 32 duplicate samples generated from 12 composite fecal sources) were analyzed by three to five core laboratories with a suite of PCR-based methods utilizing standardized reagents and protocols. Repeatability (intra-laboratory variability) and reproducibility (inter-laboratory variability) of observed results were assessed. When standardized methodologies were used, intra- and inter-laboratory %CVs were generally low (median %CV 0.1–3.3% and 1.9–7.1%, respectively) and comparable to those observed in similar inter-laboratory validation studies performed on other methods of quantifying fecal indicator bacteria (FIB) in environmental samples. ANOVA of %CV values found three human-associated methods (BsteriF1, BacHum, and HF183Taqman) to be similarly reproducible (p > 0.05) and significantly more reproducible (p < 0.05) than HumM2. This was attributed to the increased variability associated with low target concentrations detected by HumM2 (approximately 1–2 log10copies/filter lower) compared to other human-associated methods. Cow-associated methods (BacCow and CowM2) were similarly reproducible (p > 0.05). When using standardized protocols, variance component analysis indicated sample type (fecal source and concentration) to be the major contributor to total variability with that from replicate filters and inter-laboratory analysis to be within the same order of magnitude but larger than inherent intra-laboratory variability. However, when reagents and protocols were not standardized, inter-laboratory %CV generally increased with a corresponding decline in reproducibility. Overall, these findings verify the repeatability and reproducibility of these MST methods and highlight the need for standardization of protocols and consumables prior to implementation of larger scale MST studies involving multiple laboratories. |
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| title_short |
Evaluation of the repeatability and reproducibility of a suite of qPCR-based microbial source tracking methods |
| url |
https://doi.org/10.1016/j.watres.2013.01.060 |
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| author2 |
Hanley, Kaitlyn T. Cao, Yiping Badgley, Brian D. Boehm, Alexandria B. Ervin, Jared S. Goodwin, Kelly D. Gourmelon, Michèle Griffith, John F. Holden, Patricia A. Kelty, Catherine A. Lozach, Solen McGee, Charles Peed, Lindsay A. Raith, Meredith Ryu, Hodon Sadowsky, Michael J. Scott, Elizabeth A. Domingo, Jorge Santo Schriewer, Alexander Sinigalliano, Christopher D. Shanks, Orin C. Van De Werfhorst, Laurie C. Wang, Dan Wuertz, Stefan Jay, Jennifer A. |
| author2Str |
Hanley, Kaitlyn T. Cao, Yiping Badgley, Brian D. Boehm, Alexandria B. Ervin, Jared S. Goodwin, Kelly D. Gourmelon, Michèle Griffith, John F. Holden, Patricia A. Kelty, Catherine A. Lozach, Solen McGee, Charles Peed, Lindsay A. Raith, Meredith Ryu, Hodon Sadowsky, Michael J. Scott, Elizabeth A. Domingo, Jorge Santo Schriewer, Alexander Sinigalliano, Christopher D. Shanks, Orin C. Van De Werfhorst, Laurie C. Wang, Dan Wuertz, Stefan Jay, Jennifer A. |
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| doi_str |
10.1016/j.watres.2013.01.060 |
| up_date |
2024-07-06T19:59:38.707Z |
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