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[Pathogen detection in the myocardial lesion using in situ RT-PCR in mice induced by experimental CoxB(3m) virus
1Department of Cardiology Internal Medicine, Ren Ji Hospital, Shanghai Second Medical University, Shanghai 200001, China.
Objective:
To detect the location of pathogens in myocardium using in situ RT-PCR technique in order to study the pathogenetic course of the myocardial lesion induced by CoxB(3m) virus infection in mice.
Methods:
(1) Thirty and fifty Balb/c mice were used respectively to establish the acute and chronic CoxB(3m) infected models, with another 25 healthy mice as the controls; (2) KS400 image analysis system (Germany) was used to measure the cardiac chamber area and the left ventricular wall thickness of the chronic infected mice and the controls; (3) CoxB(3m) virus in myocardial tissue was detected using in situ RT-PCR by direct incorporated technique which employed nucleotide labeling by anti-digoxin antibody and bonded with alkaline phosphatase (anti-dig-AKP method).
Results:
Picture analysis indicated that the left ventricular chamber area was enlarged and the left ventricular wall was thinner in the chronic repeated virus infected models than those of the controls. With in situ RT-PCR, positive signals for Coxsackie virus B(3m) RNA were detected not only in the myocardium of the acute Balb/c mice models but also in the myocardium of the chronic mice models.
Conclusion:
Coxsackie virus B(3m) is able to induce pathologic lesions by exhibiting positive CVB-RNA signals in both acute and chronic models in mice. In the chronic experimental models, the cardiac chamber is enlarged while the ventricular wall is thinned which demonstrates the association with persistent infection of Coxsackie virus B(3m) virus.
Insights
Coxsackie virus B(3m) infection causes myocardial lesions in mice. In situ RT-PCR detected viral RNA in acute and chronic models, correlating with cardiac chamber enlargement and thinning ventricular walls.
Area of Science:
- Virology
- Pathology
- Molecular Biology
Context:
- Coxsackie virus B(3m) is a known cardiotropic virus.
- Myocardial lesions are a significant health concern.
- Understanding viral pathogenesis is crucial for developing treatments.
Purpose:
- To detect Coxsackie virus B(3m) RNA in mouse myocardium using in situ RT-PCR.
- To investigate the pathogenetic course of myocardial lesions induced by Coxsackie virus B(3m) infection.
- To correlate viral presence with cardiac structural changes.
Summary:
- Establishment of acute and chronic Coxsackie virus B(3m) infected mouse models.
- In situ RT-PCR employed to detect viral RNA in myocardial tissue.
- Image analysis revealed cardiac chamber enlargement and left ventricular wall thinning in chronic models.
Impact:
- Demonstrates Coxsackie virus B(3m) induces myocardial lesions in both acute and chronic infections.
- Highlights the utility of in situ RT-PCR for pathogen localization.
- Provides insights into the pathogenesis of Coxsackie virus B(3m)-induced cardiomyopathy.
