Development of multiplex gold nanoparticles biosensors for ultrasensitive detection and genotyping of equine herpes viruses.
Abstract: Gold nanoparticles (GNPs) biosensors can detect low viral loads and differentiate between viruses types, enabling early diagnosis and effective disease management. In the present study, we developed GNPs biosensors with two different capping agent, citrate-GNPs biosensors and polyvinylpyrrolidone (PVP)-GNPs biosensors for detection of EHV-1 and EHV-4 in multiplex real time PCR (rPCR). Citrate-GNPs and PVP-GNPs biosensors can detect dilution 10 of EHV-1 with mean Cycle threshold (Ct) 11.7 and 9.6, respectively and one copy as limit of detection, while citrate-GNPs and PVP-GNPs biosensors can detect dilution 10 of EHV-4 with mean Ct 10.5 and 9.2, respectively and one copy as limit of detection. These findings were confirmed by testing 87 different clinical samples, 4 more samples were positive with multiplex GNPs biosensors rPCR than multiplex rPCR. Multiplex citrate-GNPs and PVP-GNPs biosensors for EHV-1 and EHV-4 are a significant breakthrough in the diagnosis of these virus types. These biosensors offer high sensitivity and specificity, allowing for the accurate detection of the target viruses at very low concentrations and improve the early detection of EHV-1 and EHV-4, leading to faster control of infected animals to prevent the spread of these viruses.
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Publication Date: 2023-09-13 PubMed ID: 37704638PubMed Central: PMC10500010DOI: 10.1038/s41598-023-41918-4Google Scholar: Lookup
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Summary
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This research presents the development of gold nanoparticle biosensors for accurate detection and genotyping of equine herpes viruses (EHV-1 and EHV-4), leading to improved disease management through earlier detection and intervention.
Development of gold nanoparticle biosensors
The researchers carried out an experiment for the development of gold nanoparticle (GNPs) biosensors for the diagnosis of EHV-1 and EHV-4. The experiments involved the use of citrate-capped GNPs biosensors and polyvinylpyrrolidone-capped (PVP) GNPs biosensors for the detection of the viruses with multiplex real-time PCR (polymerase chain reaction).
- The citrate-GNPs and PVP-GNPs biosensors were able to detect the presence of EHV-1 in a ten-fold dilution with mean cycle threshold (Ct) values of 11.7 and 9.6, respectively.
- Similar results were found for EHV-4 detection, with mean Ct values of 10.5 and 9.2 for citrate-GNPs and PVP-GNPs biosensors, respectively.
- The limit of detection for both types of biosensors for both types of viruses was as low as one copy, indicating extremely high sensitivity.
Testing and validation with clinical samples
To validate the sensitivity and specificity of the developed GNPs biosensors, a set of different clinical samples were tested.
- 87 clinical samples were utilized in this testing phase.
- The testing resulted in the detection of 4 more positive samples using multiplex GNPs biosensors as compared to the traditional multiplex rPCR method.
- This clearly demonstrates the higher sensitivity of the developed GNPs biosensors.
Implication of the findings
The study signifies a breakthrough in the diagnosis of EHV-1 and EHV-4 by offering highly sensitive and specific GNPs biosensors.
- The developed biosensors have improved the potential for early detection, which is crucial for effective control and management of these diseases.
- By using the biosensors, it is possible to detect the target viruses at very low concentrations, which means that infected animals can be isolated and treated much faster, limiting the spread of the disease.
Cite This Article
APA
Ghoniem SM, ElZorkany HE, Hagag NM, El-Deeb AH, Shahein MA, Hussein HA.
(2023).
Development of multiplex gold nanoparticles biosensors for ultrasensitive detection and genotyping of equine herpes viruses.
Sci Rep, 13(1), 15140.
https://doi.org/10.1038/s41598-023-41918-4 Publication
Researcher Affiliations
- Department of Virology, Animal Health Research Institute, Agriculture Research Center, Giza, 12618, Egypt.
- Nanotechnology and Advanced Materials Central Lab, Agriculture Research Center, Giza, 12619, Egypt.
- Genome Research Unit, Animal Health Research Institute, Agriculture Research Center, Giza, 12618, Egypt.
- Department of Virology, Faculty of Veterinary Medicine, Cairo University, P.O. Box 12211, Giza, Egypt.
- Department of Virology, Faculty of Veterinary Medicine, King Salman International University, South Sinai, Egypt.
- Department of Virology, Animal Health Research Institute, Agriculture Research Center, Giza, 12618, Egypt.
- Department of Virology, Faculty of Veterinary Medicine, Cairo University, P.O. Box 12211, Giza, Egypt. husvirol@cu.edu.eg.
MeSH Terms
- Animals
- Horses
- Gold
- Genotype
- Metal Nanoparticles
- Citrates
- Citric Acid
- Herpesvirus 1, Equid / genetics
- Povidone
Conflict of Interest Statement
The authors declare no competing interests.
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