Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

66.5K
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
66.5K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

The Effect of Polyphenol Supplementation in People with Multiple Sclerosis: A Systematic Review of Clinical Trials.

Nutrients·2026
Same author

Occurrence of novel human tomato brown rugose fruit virus and conventional microbial source tracking genetic markers in a Hawaiian coupled stream-beach system.

Environmental science. Processes & impacts·2025
Same author

Novel Bacteroides bacteriophage-like quantitative real-time PCR assays for human sewage pollution measurement.

Environmental pollution (Barking, Essex : 1987)·2025
Same author

A streamlined qPCR method for characterization of Enterococcus spp. levels in ambient surface water samples.

Water research·2025
Same author

Cultured and molecular measures of fecal indicator microbes in Gulf of Mexico recreational waters.

The Science of the total environment·2025
Same author

An integrated approach to coupled nutrient and microbial source tracking in an agricultural watershed.

Water research·2024

Related Experiment Video

Updated: Mar 25, 2026

A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters
12:14

A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters

Published on: March 9, 2016

10.4K

Data Acceptance Criteria for Standardized Human-Associated Fecal Source Identification Quantitative Real-Time PCR

Orin C Shanks1, Catherine A Kelty2, Robin Oshiro3

  • 1U.S. Environmental Protection Agency, Office of Research and Development, Cincinnati, Ohio, USA shanks.orin@epa.gov.

Applied and Environmental Microbiology
|February 28, 2016
PubMed
Summary

This study establishes data acceptance criteria for human fecal source identification quantitative real-time PCR (qPCR) to improve water quality management. Standardized protocols and interlaboratory data ensure reliable results for public health protection.

More Related Videos

Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses
11:29

Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses

Published on: December 3, 2011

12.7K
Detection of Helicobacter pylori Infection and Antibiotic Resistance via Stool Quantitative Polymerase Chain Reaction Analysis
04:56

Detection of Helicobacter pylori Infection and Antibiotic Resistance via Stool Quantitative Polymerase Chain Reaction Analysis

Published on: May 16, 2025

1.4K

Related Experiment Videos

Last Updated: Mar 25, 2026

A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters
12:14

A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters

Published on: March 9, 2016

10.4K
Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses
11:29

Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses

Published on: December 3, 2011

12.7K
Detection of Helicobacter pylori Infection and Antibiotic Resistance via Stool Quantitative Polymerase Chain Reaction Analysis
04:56

Detection of Helicobacter pylori Infection and Antibiotic Resistance via Stool Quantitative Polymerase Chain Reaction Analysis

Published on: May 16, 2025

1.4K

Area of Science:

  • Environmental microbiology
  • Molecular biology
  • Water quality assessment

Background:

  • Growing interest in human fecal source identification using quantitative real-time PCR (qPCR) for water management.
  • Need for standardized protocols and data quality confidence across laboratories for qPCR applications.
  • Lack of consensus on evaluating human fecal source identification qPCR experiments hinders widespread adoption.

Purpose of the Study:

  • To establish data acceptance criteria for human fecal source identification qPCR experiments.
  • To provide benchmarks for evaluating technical quality based on interlaboratory data.
  • To facilitate the transition of qPCR from a research tool to a standardized protocol.

Main Methods:

  • Generated data from HF183/BacR287 and HumM2 human-associated qPCR methods across 14 laboratories using standardized protocols.
  • Utilized nested analysis of variance (ANOVA) to establish proficiency metrics.
  • Assessed extraneous DNA contamination and calibration model performance.

Main Results:

  • Established benchmarks for lab-to-lab variability, within-lab replicate testing, and random error.
  • Defined criteria for assessing DNA contamination and calibration model performance (correlation coefficient, amplification efficiency, lower limit of quantification).
  • Demonstrated implementation of proposed criteria with additional laboratory data.

Conclusions:

  • Proposed data acceptance criteria enhance the technical quality evaluation of human fecal source identification qPCR.
  • Standardized protocols and defined benchmarks improve confidence in water quality management data.
  • Facilitates consistent and reliable application of qPCR for public health protection.