| Development of Bacteroides 16S rRNA gene TaqMan-based real-time PCR assays for estimation of total, human, and bovine fecal pollution in water. | |
| | |
MedLine Citation:
|
PMID: 16751534 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Bacteroides species are promising indicators for differentiating livestock and human fecal contamination in water because of their high concentration in feces and potential host specificity. In this study, a real-time PCR assay was designed to target Bacteroides species (AllBac) present in human, cattle, and equine feces. Direct PCR amplification (without DNA extraction) using the AllBac assay was tested on feces diluted in water. Fecal concentrations and threshold cycle were linearly correlated, indicating that the AllBac assay can be used to estimate the total amount of fecal contamination in water. Real-time PCR assays were also designed for bovine-associated (BoBac) and human-associated (HuBac) Bacteroides 16S rRNA genes. Assay specificities were tested using human, bovine, swine, canine, and equine fecal samples. The BoBac assay was specific for bovine fecal samples (100% true-positive identification; 0% false-positive identification). The HuBac assay had a 100% true-positive identification, but it also had a 32% false-positive rate with potential for cross-amplification with swine feces. The assays were tested using creek water samples from three different watersheds. Creek water did not inhibit PCR, and results from the AllBac assay were correlated with those from Escherichia coli concentrations (r2= 0.85). The percentage of feces attributable to bovine and human sources was determined for each sample by comparing the values obtained from the BoBac and HuBac assays with that from the AllBac assay. These results suggest that real-time PCR assays without DNA extraction can be used to quantify fecal concentrations and provide preliminary fecal source identification in watersheds. |
| | |
Authors:
|
Alice Layton; Larry McKay; Dan Williams; Victoria Garrett; Randall Gentry; Gary Sayler |
Related Documents
:
|
15862454 - Quantification of staphylococcus aureus in unpasteurised bovine and caprine milk by rea... 18623994 - Rapid and sensitive detection of ms2 coliphages in wastewater samples by quantitative r... 19083844 - Interrogation of short tandem repeats using fluorescent probes and melting curve analys... 17691694 - Comparison of real-time polymerase chain reaction and conventional polymerase chain rea... 12841954 - A sensitive and specific method for the determination of total ribavirin in human red b... 18092754 - Dose formulation and analysis of diapocynin. |
Publication Detail:
|
Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
|
Title: Applied and environmental microbiology Volume: 72 ISSN: 0099-2240 ISO Abbreviation: Appl. Environ. Microbiol. Publication Date: 2006 Jun |
Date Detail:
|
Created Date: 2006-06-05 Completed Date: 2006-08-30 Revised Date: 2009-11-18 |
Medline Journal Info:
|
Nlm Unique ID: 7605801 Medline TA: Appl Environ Microbiol Country: United States |
Other Details:
|
Languages: eng Pagination: 4214-24 Citation Subset: IM |
Affiliation:
|
The University of Tennessee, Center for Environmental Biotechnology, 676 Dabney Hall, Knoxville, TN 37996-1605, USA. alayton@utk.edu |
| Data Bank Information | |
Bank Name/Acc. No.:
|
GENBANK/AY597127; AY597128; AY597129; AY597130; AY597131; AY597132; AY597133; AY597134; AY597135; AY597136; AY597137; AY597138; AY597139; AY597140; AY597141; AY597142; AY597143; AY597144; AY597145; AY597146; AY597147; AY597148; AY597149; AY597150; AY597151; AY597152; AY597153; AY597154; AY597155; AY597156; AY597157; AY597158; AY597159; AY597160; AY597161; AY597162; AY597163; AY597164; AY597165; AY597166; AY597167; AY597168; AY597169; AY597170; AY597171; AY597172; AY597173; AY597174; AY597175; AY597176; AY597177; AY597178; AY597179; AY597180; AY597181; AY597182; AY597183; AY597184; AY597185; AY597186; AY597187; AY597188; AY597189; AY597190; AY597191; AY597192; AY597193; AY597194; AY597195; AY597196; AY597197; AY597198; AY597199; AY597200; AY597201; AY597202; AY597203; AY597204; AY597205; AY597206 |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Bacteroides / classification, genetics* Base Sequence Cattle DNA Primers DNA, Bacterial / genetics DNA, Ribosomal / genetics Feces / microbiology* Humans Molecular Sequence Data Phylogeny Polymerase Chain Reaction / methods* RNA, Bacterial / genetics RNA, Ribosomal, 16S / genetics* Water Pollution* |
| Chemical | |
Reg. No./Substance:
|
0/DNA Primers; 0/DNA, Bacterial; 0/DNA, Ribosomal; 0/RNA, Bacterial; 0/RNA, Ribosomal, 16S |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Production of xylitol from D-xylose by a xylitol dehydrogenase gene-disrupted mutant of Candida trop...
Next Document: Dipeptide synthesis by an aminopeptidase from Streptomyces septatus TH-2 and its application to synt...