Development and validation of rt-qpcr for vesicular stomatitis virus detection (Alagoas vesiculovirus).

The research aimed to develop a technique for quicker and more accurate detection of vesicular stomatitis virus, an infectious disease often found in Western Hemisphere countries and affecting livestock. The scientists carried out validation tests on a multiplex RT-qPCR method for identifying RNA from Alagoas vesiculovirus, and found it demonstrated high sensitivity, specificity, repeatability, and reproducibility.

Objective of the Research

• The focus was on Vesicular stomatitis, an infectious animal disease widespread in Western Hemisphere countries, affecting creatures such as cattle, swine, and horses.
• Given that the virus’ symptoms in these animals are closely similar to the foot-and-mouth disease, a very rapid and correct differential diagnosis is a major requirement for managing the infection effectively.
• The main goal of this research was to create and perform validation tests for multiplex RT-qPCR, a method used for detecting the RNA of the Alagoas vesiculovirus with improved accuracy and efficiency.

Methods and Test Parameters

• The team tested the multiplex RT-qPCR technique based on multiple parameters including sensitivity, analytical specificity, analytical performance, repeatability, reproducibility and the uncertainty of measurement.
• The threshold cycle values noted from each experimental sample, done in triplicate, were evaluated by considering any variations occurring between days, analysts, and equipment.
• An analysis of variance was performed to discern the variances of repeatability and reproducibility.

Research Findings

• Findings from the validation tests showed that RT-qPCRs (Real-Time Quantitative Reverse Transcription PCR) had excellent sensitivity and specificity in detecting the RNA of Alagoas vesiculovirus.
• The examined validation parameters had low coefficients of variation, indicating excellent repeatability and reproducibility.
• The results obtained were in line with those found in other validation studies, proving their validity and precision in identifying the Vesicular stomatitis virus.

Significance of the Findings

• A reliable and efficient detection method like the RT-qPCR for this disease can greatly aid in carrying out appropriate preventive measures in a timely manner.
• Given that this infectious disease affects major livestock and exhibits symptoms similar to another major animal disease, accurate differential diagnosis can prevent economic losses and large-scale spread of the disease.
• A validated detection method will not only benefit livestock health, but will also impact the economic health of farmers, animal industry stakeholders, and potentially affect public health in certain cases.