Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study
The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contr...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Schriewer, Alexander [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: |
12 |
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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:6909-6920 ; extent:12 |
| Links: |
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| DOI / URN: |
10.1016/j.watres.2013.03.062 |
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ELV021577757 |
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| 520 | |a The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. | ||
| 520 | |a The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. | ||
| 650 | 7 | |a Canine fecal pollution |2 Elsevier | |
| 650 | 7 | |a Bacteroidales |2 Elsevier | |
| 650 | 7 | |a Microbial source tracking |2 Elsevier | |
| 650 | 7 | |a Quantitative PCR |2 Elsevier | |
| 700 | 1 | |a Goodwin, Kelly D. |4 oth | |
| 700 | 1 | |a Sinigalliano, Christopher D. |4 oth | |
| 700 | 1 | |a Cox, Annie M. |4 oth | |
| 700 | 1 | |a Wanless, David |4 oth | |
| 700 | 1 | |a Bartkowiak, Jakob |4 oth | |
| 700 | 1 | |a Ebentier, Darcy L. |4 oth | |
| 700 | 1 | |a Hanley, Kaitlyn T. |4 oth | |
| 700 | 1 | |a Ervin, Jared |4 oth | |
| 700 | 1 | |a Deering, Louise A. |4 oth | |
| 700 | 1 | |a Shanks, Orin C. |4 oth | |
| 700 | 1 | |a Peed, Lindsay A. |4 oth | |
| 700 | 1 | |a Meijer, Wim G. |4 oth | |
| 700 | 1 | |a Griffith, John F. |4 oth | |
| 700 | 1 | |a SantoDomingo, Jorge |4 oth | |
| 700 | 1 | |a Jay, Jennifer A. |4 oth | |
| 700 | 1 | |a Holden, Patricia A. |4 oth | |
| 700 | 1 | |a Wuertz, Stefan |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 |
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10.1016/j.watres.2013.03.062 doi GBVA2013001000018.pica (DE-627)ELV021577757 (ELSEVIER)S0043-1354(13)00547-2 DE-627 ger DE-627 rakwb eng 550 550 DE-600 333.7 320 VZ Schriewer, Alexander verfasserin aut Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study 2013transfer abstract 12 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. Canine fecal pollution Elsevier Bacteroidales Elsevier Microbial source tracking Elsevier Quantitative PCR Elsevier Goodwin, Kelly D. oth Sinigalliano, Christopher D. oth Cox, Annie M. oth Wanless, David oth Bartkowiak, Jakob oth Ebentier, Darcy L. oth Hanley, Kaitlyn T. oth Ervin, Jared oth Deering, Louise A. oth Shanks, Orin C. oth Peed, Lindsay A. oth Meijer, Wim G. oth Griffith, John F. oth SantoDomingo, Jorge oth Jay, Jennifer A. oth Holden, Patricia A. oth Wuertz, Stefan 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:6909-6920 extent:12 https://doi.org/10.1016/j.watres.2013.03.062 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U AR 47 2013 18 15 1115 6909-6920 12 045F 550 |
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10.1016/j.watres.2013.03.062 doi GBVA2013001000018.pica (DE-627)ELV021577757 (ELSEVIER)S0043-1354(13)00547-2 DE-627 ger DE-627 rakwb eng 550 550 DE-600 333.7 320 VZ Schriewer, Alexander verfasserin aut Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study 2013transfer abstract 12 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. Canine fecal pollution Elsevier Bacteroidales Elsevier Microbial source tracking Elsevier Quantitative PCR Elsevier Goodwin, Kelly D. oth Sinigalliano, Christopher D. oth Cox, Annie M. oth Wanless, David oth Bartkowiak, Jakob oth Ebentier, Darcy L. oth Hanley, Kaitlyn T. oth Ervin, Jared oth Deering, Louise A. oth Shanks, Orin C. oth Peed, Lindsay A. oth Meijer, Wim G. oth Griffith, John F. oth SantoDomingo, Jorge oth Jay, Jennifer A. oth Holden, Patricia A. oth Wuertz, Stefan 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:6909-6920 extent:12 https://doi.org/10.1016/j.watres.2013.03.062 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U AR 47 2013 18 15 1115 6909-6920 12 045F 550 |
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10.1016/j.watres.2013.03.062 doi GBVA2013001000018.pica (DE-627)ELV021577757 (ELSEVIER)S0043-1354(13)00547-2 DE-627 ger DE-627 rakwb eng 550 550 DE-600 333.7 320 VZ Schriewer, Alexander verfasserin aut Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study 2013transfer abstract 12 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. Canine fecal pollution Elsevier Bacteroidales Elsevier Microbial source tracking Elsevier Quantitative PCR Elsevier Goodwin, Kelly D. oth Sinigalliano, Christopher D. oth Cox, Annie M. oth Wanless, David oth Bartkowiak, Jakob oth Ebentier, Darcy L. oth Hanley, Kaitlyn T. oth Ervin, Jared oth Deering, Louise A. oth Shanks, Orin C. oth Peed, Lindsay A. oth Meijer, Wim G. oth Griffith, John F. oth SantoDomingo, Jorge oth Jay, Jennifer A. oth Holden, Patricia A. oth Wuertz, Stefan 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:6909-6920 extent:12 https://doi.org/10.1016/j.watres.2013.03.062 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U AR 47 2013 18 15 1115 6909-6920 12 045F 550 |
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10.1016/j.watres.2013.03.062 doi GBVA2013001000018.pica (DE-627)ELV021577757 (ELSEVIER)S0043-1354(13)00547-2 DE-627 ger DE-627 rakwb eng 550 550 DE-600 333.7 320 VZ Schriewer, Alexander verfasserin aut Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study 2013transfer abstract 12 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. Canine fecal pollution Elsevier Bacteroidales Elsevier Microbial source tracking Elsevier Quantitative PCR Elsevier Goodwin, Kelly D. oth Sinigalliano, Christopher D. oth Cox, Annie M. oth Wanless, David oth Bartkowiak, Jakob oth Ebentier, Darcy L. oth Hanley, Kaitlyn T. oth Ervin, Jared oth Deering, Louise A. oth Shanks, Orin C. oth Peed, Lindsay A. oth Meijer, Wim G. oth Griffith, John F. oth SantoDomingo, Jorge oth Jay, Jennifer A. oth Holden, Patricia A. oth Wuertz, Stefan 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:6909-6920 extent:12 https://doi.org/10.1016/j.watres.2013.03.062 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U AR 47 2013 18 15 1115 6909-6920 12 045F 550 |
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10.1016/j.watres.2013.03.062 doi GBVA2013001000018.pica (DE-627)ELV021577757 (ELSEVIER)S0043-1354(13)00547-2 DE-627 ger DE-627 rakwb eng 550 550 DE-600 333.7 320 VZ Schriewer, Alexander verfasserin aut Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study 2013transfer abstract 12 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. Canine fecal pollution Elsevier Bacteroidales Elsevier Microbial source tracking Elsevier Quantitative PCR Elsevier Goodwin, Kelly D. oth Sinigalliano, Christopher D. oth Cox, Annie M. oth Wanless, David oth Bartkowiak, Jakob oth Ebentier, Darcy L. oth Hanley, Kaitlyn T. oth Ervin, Jared oth Deering, Louise A. oth Shanks, Orin C. oth Peed, Lindsay A. oth Meijer, Wim G. oth Griffith, John F. oth SantoDomingo, Jorge oth Jay, Jennifer A. oth Holden, Patricia A. oth Wuertz, Stefan 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:6909-6920 extent:12 https://doi.org/10.1016/j.watres.2013.03.062 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U AR 47 2013 18 15 1115 6909-6920 12 045F 550 |
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Enthalten in Matches, mismatches and priorities of pathways from a climate-resilient development perspective in the mountains of Nepal Amsterdam [u.a.] volume:47 year:2013 number:18 day:15 month:11 pages:6909-6920 extent:12 |
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Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study |
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The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. |
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The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. |
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The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment. |
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The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment.</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment.</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">Canine fecal pollution</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">Bacteroidales</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">Microbial source tracking</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="650" ind1=" " ind2="7"><subfield code="a">Quantitative PCR</subfield><subfield code="2">Elsevier</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Goodwin, Kelly D.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Sinigalliano, Christopher D.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Cox, Annie M.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Wanless, David</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Bartkowiak, Jakob</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Ebentier, Darcy L.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Hanley, Kaitlyn T.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Ervin, Jared</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Deering, Louise A.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Shanks, Orin C.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Peed, Lindsay A.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Meijer, Wim G.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Griffith, John F.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">SantoDomingo, Jorge</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Jay, Jennifer A.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Holden, Patricia A.</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Wuertz, Stefan</subfield><subfield code="4">oth</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="i">Enthalten in</subfield><subfield code="n">Elsevier Science</subfield><subfield code="a">Pandey, Avash ELSEVIER</subfield><subfield code="t">Matches, mismatches and priorities of pathways from a climate-resilient development perspective in the mountains of Nepal</subfield><subfield code="d">2021</subfield><subfield code="d">a journal of the International Association on Water Quality (IAWQ)</subfield><subfield code="g">Amsterdam [u.a.]</subfield><subfield code="w">(DE-627)ELV006716016</subfield></datafield><datafield tag="773" ind1="1" ind2="8"><subfield code="g">volume:47</subfield><subfield code="g">year:2013</subfield><subfield code="g">number:18</subfield><subfield code="g">day:15</subfield><subfield code="g">month:11</subfield><subfield code="g">pages:6909-6920</subfield><subfield code="g">extent:12</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">https://doi.org/10.1016/j.watres.2013.03.062</subfield><subfield code="3">Volltext</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_USEFLAG_U</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_ELV</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">SYSFLAG_U</subfield></datafield><datafield tag="951" ind1=" " ind2=" "><subfield code="a">AR</subfield></datafield><datafield tag="952" ind1=" " ind2=" "><subfield code="d">47</subfield><subfield code="j">2013</subfield><subfield code="e">18</subfield><subfield code="b">15</subfield><subfield code="c">1115</subfield><subfield code="h">6909-6920</subfield><subfield code="g">12</subfield></datafield><datafield tag="953" ind1=" " ind2=" "><subfield code="2">045F</subfield><subfield code="a">550</subfield></datafield></record></collection>
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