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Novel Nucleic Acid Detection for Human Parvovirus B19 Based on Pyrococcus furiosus Argonaute Protein
Weiran Chen1, Liyang Qiu1, Ting Luo1
1State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Key Laboratory of Industrial Biotechnology, Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, School of Life Sciences, Hubei University, Wuhan 430062, China.
Viruses
|March 30, 2023
Summary
A new Parvovirus B19 (B19V) diagnostic tool using Pyrococcus furiosus Argonaute (PfAgo) offers high sensitivity. This method shows promise for accurate B19V infection detection and epidemiological studies.
Area of Science:
- Molecular Biology
- Biosensing Technology
- Virology
Background:
- Parvovirus B19 (B19V) causes human diseases, yet no vaccines or antiviral treatments exist.
- Accurate diagnosis of B19V infection is crucial due to its pathogenicity.
- Previous Clustered Regularly Interspaced Palindromic Repeats (CRISPR)-Cas12a (cpf1)-based electrochemical biosensor (E-CRISPR) achieved picomole sensitivity.
Purpose of the Study:
- To develop a novel nucleic acid detection system for B19V using Pyrococcus furiosus Argonaute (PfAgo).
- To target the nonstructural protein 1 (NS1) region of the B19V genome for detection.
- To evaluate the sensitivity and diagnostic accuracy of the PfAgo-based system.
Main Methods:
- Established a novel nucleic acid detection system based on Pyrococcus furiosus Argonaute (PfAgo).
- Targeted the nonstructural protein 1 (NS1) region of the B19V viral genome (B19-NS1 PAND).
- Assessed Minimum Detectable Concentration (MDC) with and without Polymerase Chain Reaction (PCR) amplification.
Main Results:
- Without preamplification, B19-NS1 PAND achieved a Minimum Detectable Concentration (MDC) of approximately 4 nM.
- With amplification, the MDC was reduced to the attomolar (aM) level (54 aM), significantly surpassing E-CRISPR sensitivity.
- Diagnostic results from clinical samples showed 100% consistency with PCR and Sanger sequencing.
Conclusions:
- The PfAgo-mediated nucleic acid detection system (B19-NS1 PAND) demonstrates high sensitivity for B19V detection.
- This system, especially with amplification, offers a sensitive alternative to existing diagnostic methods.
- B19-NS1 PAND shows potential for molecular testing in clinical diagnosis and B19V epidemiological investigations.
