Oct 17, 2023

Public workspaceTroubleshooting guide for DDNS

  • 1Imperial College London;
  • 2Medicines and Healthcare products Regulatory Agency;
  • 3University of Edinburgh
Open access
Document CitationJoyce Akello, Alex Shaw, Catherine Troman, Erika Bujaki, Manasi Majumdar, c.ansley, Zoe Vance, rachel.colquhoun, Andrew Rambaut, Javier Martin, Nick Grassly, Aine OToole 2023. Troubleshooting guide for DDNS. protocols.io https://dx.doi.org/10.17504/protocols.io.yxmvm3w25l3p/v1
License: This is an open access document distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Created: October 17, 2023
Last Modified: April 19, 2024
Document Integer ID: 89412
Funders Acknowledgement:
Bill and Melinda Gates Foundation
Abstract
This document provides general guidance for troubleshooting problems encountered in the DDNS method. It outlines the factors that might influence the outcome of the DDNS PCR assay run, the common problems encountered, possible causes of the problems, and provides recommendations on corrective action and preventative action.
This document is primarily developed for DDNS performed on stool samples, but included are other factors to consider for other sample types.
Purpose
To outline the most common sources of DDNS assay run failure / unacceptable results, reduced/lower sensitivity
than expected and provide recommendation on how to avoid these pitfalls and
improve performance.
Associated Forms & Documents
DDNS Protocol v2.0 (protocols.io)
Quality Control and Data Recording for DDNS (protocols.io)
Poliovirus detection and nanopore sequencing FAQs (Poliovirus detection and nanopore sequencing
Verification guidelines for the DDNS method for poliovirus direct detection (protocols.io)

Troubleshooting considerations.

Table1. Factors to consider when troubleshooting DDNS results.


Table 2. Commonly encountered DDNS PCR assay run errors, possible causes, recommended corrective, and preventive actions.


Table 3: Commonly encountered errors when installing and running PIRANHA GUI