Short RNA indicator sequences are not completely degraded by autoclaving

Unnithan, Veena V.; Unc, Adrian; Joe, Valerisa; Smith, Geoffrey B.

doi:10.1038/srep04070

Cited by 8 publications

(5 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The reagent used for decontamination of equipment and surfaces is critical to maintaining clean standards. For example, standard autoclaving is inadequate for destroying nucleic acids (Unnithan et al 2014). Of the generally applied decontaminating solutions, sodium hypochlorite (bleach) is the most effective for removing DNA and PCR products (Prince & Andrus 1992;Champlot et al 2010).…”

Section: Clean Practices In the Laboratorymentioning

confidence: 99%

Critical considerations for the application of environmental DNA methods to detect aquatic species

et al. 2016

View full text Add to dashboard Cite

Summary1. Species detection using environmental DNA (eDNA) has tremendous potential for contributing to the understanding of the ecology and conservation of aquatic species. Detecting species using eDNA methods, rather than directly sampling the organisms, can reduce impacts on sensitive species and increase the power of field surveys for rare and elusive species. The sensitivity of eDNA methods, however, requires a heightened awareness and attention to quality assurance and quality control protocols. Additionally, the interpretation of eDNA data demands careful consideration of multiple factors. As eDNA methods have grown in application, diverse approaches have been implemented to address these issues. With interest in eDNA continuing to expand, supportive guidelines for undertaking eDNA studies are greatly needed. 2. Environmental DNA researchers from around the world have collaborated to produce this set of guidelines and considerations for implementing eDNA methods to detect aquatic macroorganisms. 3. Critical considerations for study design include preventing contamination in the field and the laboratory, choosing appropriate sample analysis methods, validating assays, testing for sample inhibition and following minimum reporting guidelines. Critical considerations for inference include temporal and spatial processes, limits of correlation of eDNA with abundance, uncertainty of positive and negative results, and potential sources of allochthonous DNA. 4. We present a synthesis of knowledge at this stage for application of this new and powerful detection method.

show abstract

Section: Clean Practices In the Laboratorymentioning

confidence: 99%

Critical considerations for the application of environmental DNA methods to detect aquatic species

et al. 2016

View full text Add to dashboard Cite

show abstract

“…This clearly indicated non-specific amplification. Since this was not true for the shorter ''A'' amplicons, this suggests differential degradation of various RNA sequences [14]. Thus, the treatments here were evaluated using a very stable indicator sequence [14].…”

Section: Resultsmentioning

confidence: 99%

Enzymatic Pre-treatment of Wastewater to Minimize Recovery by Reverse Transcriptase PCR of RNA from Inactive Bacteriophages

2015

Self Cite

View full text Add to dashboard Cite

Quantitative viral risk assessments for wastewaters are notoriously difficult. The often considered quantitative reverse transcriptase PCR reflects poorly on virus infectivity rates leading to inaccurate risk interpretations. Various techniques focused on the degradation of the nucleic acids of non-infective viruses were previously employed. We comparatively assessed the effectiveness of such enzymatic treatments for MS2 bacteriophage in treated wastewaters. The single use of RNase A at an appropriate concentration may be as effective as the combination of RNase followed by Proteinase K and more rapid. While all tested enzymatic treatments minimized recovery of RNA (>95 %) in the absence of infective MS2, none completely eliminated the signal recovery. Selection of any enzymatic protocol for minimizing recovery of RNA from degraded, non-infective viruses should balance the methods efficacy with its expediency.

show abstract

“…However, the maintenance time of autoclaving at 121 °C is generally about 30 min, which is not enough to completely degrade the nucleic acid of pathogenic microorganisms. Studies have shown that after autoclaving, shorter DNA or RNA fragments produced by incomplete degradation of viral nucleic acid may be recovered by molecular amplification techniques (Choi et al 2014 ; Unnithan et al 2014 ). Our results also showed that autoclaving could not completely degrade the viral nucleic acid.…”

Section: Discussionmentioning

confidence: 99%

Investigation of potential safety hazards during medical waste disposal in SARS-CoV-2 testing laboratory

Jun

Jin

Xue

et al. 2021

Environ Sci Pollut Res

View full text Add to dashboard Cite

This study aims to investigate the potential safety hazards and provide reference for improving the medical waste disposal procedure in SARS-CoV-2 testing laboratory. Our SARS-CoV-2 testing group detected the RNA residue on the surface of medical waste with Droplet Digital PCR, and held a meeting to discuss the risks in the laboratory medical waste disposal process. After effective autoclaving, SARS-CoV-2 contaminated on the surface of medical waste bags was killed, but the average concentration of viral RNA residues was still 0.85 copies/cm2. It would not pose a health risk, but might contaminate the laboratory and affect the test results. When the sterilized medical waste bags were transferred directly by the operators without hand disinfection, re-contamination would happen, which might cause the virus to leak out of the laboratory. Furthermore, we found that sterilization effect monitoring and cooperation among operators were also very important. In summary, we investigated and analyzed the potential safety hazards during the medical waste disposal process in SARS-CoV-2 testing laboratory, and provided reasonable suggestions to ensure the safety of medical waste disposal.

show abstract

Short RNA indicator sequences are not completely degraded by autoclaving

Cited by 8 publications

References 20 publications

Critical considerations for the application of environmental DNA methods to detect aquatic species

Critical considerations for the application of environmental DNA methods to detect aquatic species

Enzymatic Pre-treatment of Wastewater to Minimize Recovery by Reverse Transcriptase PCR of RNA from Inactive Bacteriophages

Investigation of potential safety hazards during medical waste disposal in SARS-CoV-2 testing laboratory

Contact Info

Product

Resources

About