Development of a fluorescent probe-based recombinase polymerase amplification assay for rapid detection of Orf virus.

Virol J

State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.

Published: December 2015

Background: Orf virus (ORFV) is the causative agent of Orf (also known as contagious ecthyma or contagious papular dermatitis), a severe infectious skin disease in goats, sheep and other ruminants. The rapid detection of ORFV is of great importance in disease control and highly needed. A isothermal molecular diagnostic approach, termed recombinase polymerase amplification (RPA), is considered as an novel and rapid alternative techonology to PCR assay.

Results: In the present study, a novel fluorescent probe based on RPA assay (ORFV exo RPA assay) was developed. The developed ORFV exo RPA assay was capable of as low as 100 copies of ORFV DNA /reaction and was highly specific, with no cross-reaction with closely related viruses (capripox virus, foot-and-mouth disease virus or peste des petits ruminants virus). Further assessment with clinical samples showed that the developed ORFV exo RPA assay has good correlation with qPCR assays for detection of ORFV.

Conclusions: These results suggest that the developed ORFV exo RPA assay is suitable for rapid detection of ORFV.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4668657PMC
http://dx.doi.org/10.1186/s12985-015-0440-zDOI Listing

Publication Analysis

Top Keywords

rpa assay
20
orfv exo
16
exo rpa
16
rapid detection
12
developed orfv
12
recombinase polymerase
8
polymerase amplification
8
orf virus
8
orfv
8
detection orfv
8

Similar Publications

Tuberculosis (TB) is the second deadliest infectious disease worldwide. Current TB diagnostics utilize sputum samples, which are difficult to obtain, and sample processing is time-consuming and difficult. This study developed an integrated diagnostic platform for the rapid visual detection of Mycobacterium tuberculosis (Mtb) in breath samples at the point-of-care (POC), especially in resource-limited settings.

View Article and Find Full Text PDF

Short-Time Preamplification-Assisted One-Pot CRISPR Nucleic Acid Detection Method with Portable Self-Heating Equipment for Point-of-Care Diagnosis.

Anal Chem

January 2025

State Key Laboratory for Manufacturing Systems Engineering, School of Instrument Science and Techonology, Xi'an Jiaotong University, Xi'an 710054, China.

Infectious diseases, especially respiratory infections, have been significant threats to human health. Therefore, it is essential to develop rapid, portable, and highly sensitive diagnostic methods for their control. Herein, a short-time preamplified, one-pot clustered regularly interspaced short palindromic repeats (CRISPR) nucleic acid detection method (SPOC) is developed by combining the rapid recombinase polymerase amplification (RPA) with CRISPR-Cas12a to reduce the mutual interference and achieve facile and rapid molecular diagnosis.

View Article and Find Full Text PDF

Rapid and accurate molecular diagnostics are crucial for preventing the global spread of emerging infectious diseases. However, the current gold standard for nucleic acid detection, reverse transcription polymerase chain reaction (RT-PCR), relies heavily on traditional magnetic beads or silica membranes for nucleic acid extraction, resulting in several limitations, including time-consuming processes, the need for trained personnel, and complex equipment. Therefore, there is an urgent need for fully integrated nucleic acid detection technologies that are simple to operate, rapid, and highly sensitive to meet unmet clinical needs.

View Article and Find Full Text PDF

Unlabelled: Cardamom mosaic virus causing mosaic/ disease is the most destructive virus infecting cardamom. The development of effective diagnostic assays is essential for the production of virus-free plants, as the primary spread of the virus occurs through vegetative propagation. Currently used PCR-based assays are not suitable for Point-of-Care testing, require sophisticated equipment, and are time-consuming.

View Article and Find Full Text PDF

Ligation-recognition triggered RPA-Cas12a cis-cleavage fluorogenic RNA aptamer for one-pot and label-free detection of MicroRNA in breast cancer.

Biosens Bioelectron

December 2024

Key Laboratory for Green Organic Synthesis and Application of Hunan Province, Key Laboratory of Environmentally Friendly Chemistry and Application of Ministry of Education, College of Chemistry, Xiangtan University, Xiangtan, 411105, China. Electronic address:

Article Synopsis
  • A novel one-pot assay called LRPA-CRISPR is developed for detecting miRNA, combining the efficiency of RPA and CRISPR/Cas12a systems.
  • The assay amplifies specific miRNA sequences and utilizes a cis-cleavage mechanism to produce a fluorescent signal, enabling rapid detection within 40 minutes.
  • This method boasts high sensitivity, label-free detection, and potential application in breast cancer biomarker diagnostics, making it a versatile tool for research and clinical settings.
View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!