Related Experiment Video
Updated: Jul 28, 2025
EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR
Published on: January 16, 2016
Qualitative detection of enterovirus D68 from PrimeStore® molecular transport medium: implications for home- and
Hai Nguyen-Tran1, Samantha Reno2, Eric Mwangi2
1Department of Pediatrics, Section of Infectious Diseases, University of Colorado School of Medicine, Aurora, CO, USA.
Abstract:
To ensure proper specimen handling for detecting pathogens, like Enterovirus D68 (EV-D68), from home- and self-collection, alternative techniques are needed to ensure safe transport and reliable testing. PrimeStore® Molecular Transport Medium (MTM) may be an option since it does not require cold storage and inactivates virus while preserving RNA for detection. The purpose of this validation study was to demonstrate the ability to detect EV-D68 via rRT-PCR in MTM. Using a quantified EV-D68 positive control standard, MTM limit of detection for EV-D68 RNA is 104 cp/mL and RNA remains stable up to 30 days unfrozen. Positive and negative residual respiratory specimens from the 2018 EV-D68 outbreak were used for clinical testing. There was an 80% positive and 100% negative agreement with samples in MTM compared to reference. This study demonstrates the feasibility of EV-D68 detection from respiratory specimens collected and stored in PrimeStore® MTM, with implications for home- and self-collection.
Insights
PrimeStore® Molecular Transport Medium (MTM) enables reliable detection of Enterovirus D68 (EV-D68) from respiratory specimens. This method supports home-based collection and testing without cold storage, improving pathogen surveillance.
Area of Science:
- Virology
- Molecular Diagnostics
- Public Health
Background:
- Accurate detection of respiratory pathogens like Enterovirus D68 (EV-D68) is crucial for public health surveillance.
- Home- and self-collection methods require validated transport media for reliable diagnostic testing.
- Traditional specimen transport often necessitates cold chain maintenance, posing logistical challenges.
Purpose of the Study:
- To validate the use of PrimeStore® Molecular Transport Medium (MTM) for the detection of EV-D68 RNA.
- To assess the stability of EV-D68 RNA in MTM under ambient conditions.
- To evaluate the performance of MTM for EV-D68 detection in clinical respiratory specimens.
Main Methods:
- Quantified EV-D68 positive control standards were used to determine the limit of detection (LOD) in MTM.
- EV-D68 RNA stability was assessed over 30 days at room temperature.
- Residual respiratory specimens from the 2018 EV-D68 outbreak were tested for EV-D68 using real-time reverse transcription polymerase chain reaction (rRT-PCR) after storage in MTM.
Main Results:
- The limit of detection for EV-D68 RNA in MTM was determined to be 10^4 cp/mL.
- EV-D68 RNA remained stable in MTM for up to 30 days without refrigeration.
- Clinical testing showed 80% positive agreement and 100% negative agreement compared to reference methods.
Conclusions:
- PrimeStore® MTM is a feasible medium for the detection of EV-D68 from respiratory specimens.
- MTM facilitates reliable molecular testing of EV-D68, supporting ambient temperature transport.
- This approach has significant implications for enabling widespread home- and self-collection strategies for infectious disease surveillance.
