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The M and N genes-based simplex and multiplex PCRs are better than the F or H gene-based simplex PCR for
A George1, P Dhar, B P Sreenivasa
1Division of Virology, Indian Veterinary Research Institute, Mukteswar, Nainital, India.
Abstract:
Nucleocapsid (N), matrix (M) and hemagglutinin (H) genes-based simplex PCRs and an N and M genes-based multiplex PCR were developed for detection of Peste-des-petits-ruminants virus (PPRV). The M gene PCR was the most sensitive, followed by N, H and an already described fusion (F) gene PCRs, as they could detect the virus in samples with titers of 101, 102, 104and 105 TCID50/ml, respectively. The multiplex PCR was as sensitive as the M gene PCR, but it had the advantage of differentiating PPRV from Rinderpest virus (RPV).
Insights
This study developed new PCR methods for detecting Peste-des-petits-ruminants virus (PPRV). The matrix (M) gene PCR and a multiplex PCR showed the highest sensitivity for PPRV detection and differentiation.
Area of Science:
- Veterinary Virology
- Molecular Diagnostics
- Ovine and Caprine Diseases
Background:
- Peste-des-petits-ruminants virus (PPRV) causes significant economic losses in small ruminants globally.
- Accurate and sensitive diagnostic tools are crucial for controlling PPRV outbreaks.
- Existing diagnostic methods may vary in sensitivity and specificity.
Purpose of the Study:
- To develop and evaluate novel simplex and multiplex PCR assays for the sensitive detection of PPRV.
- To compare the sensitivity of PCR assays targeting different PPRV genes (N, M, H, F).
- To assess the ability of a multiplex PCR to differentiate PPRV from Rinderpest virus (RPV).
Main Methods:
- Development of simplex PCR assays targeting Nucleocapsid (N), Matrix (M), and Hemagglutinin (H) genes of PPRV.
- Development of a multiplex PCR assay targeting N and M genes of PPRV.
- Sensitivity testing using serial dilutions of PPRV samples (TCID50/ml).
- Evaluation of the multiplex PCR's ability to differentiate PPRV from RPV.
Main Results:
- The M gene-based simplex PCR demonstrated the highest sensitivity, detecting PPRV at titers of 10^1 TCID50/ml.
- The N and H gene simplex PCRs and the previously described F gene PCR showed sensitivities of 10^2, 10^4, and 10^5 TCID50/ml, respectively.
- The multiplex PCR was as sensitive as the M gene simplex PCR and could distinguish PPRV from RPV.
Conclusions:
- The M gene simplex PCR and the N and M genes-based multiplex PCR are highly sensitive methods for PPRV detection.
- The developed multiplex PCR offers a valuable tool for simultaneous detection and differentiation of PPRV and RPV, aiding in disease control strategies.
