Acta Scientific Veterinary Sciences (ISSN: 2582-3183)

Research Article Volume 5 Issue 2

Optimization and Development of Protocol for Detection of Foot-And-Mouth Disease Virus by Negative Staining Using Transmission Electron Microscopy

Rajeev Ranjan*, Jitendra Kumar Biswal and Rabindra Prasad Singh

ICAR-Directorate of Foot and Mouth Disease, International Centre for Foot and Mouth Disease, Arugul, Bhubaneswar, Odisha, India

*Corresponding Author: Rajeev Ranjan, ICAR-Directorate of Foot and Mouth Disease, International Centre for Foot and Mouth Disease, Arugul, Bhubaneswar, Odisha, India.

Received: December 08, 2022; Published: January 09, 2023

Abstract

Using transmission electron microscopy (TEM), a specimen can be ready for examination within 10 minutes, and an experienced virologist, pathologist, or technologist can identify a pathogen morphologically. There is paucity of literature with respect to study of TEM as a tool for the detection of foot-and-mouth disease (FMD) virus in suspension in diagnostic pathology. The current study aimed to optimize and develop the process of detection of foot-and-mouth disease virus and its capsid in TEM by negative staining methods. FMD virus infected BHK-21 cell and mock infected cells were centrifuged for 5 minutes at 12000 rpm, and the supernatant was used for negative staining. The supernatant was placed on the TEM grid using either single droplet touch methods or direct application of a single drop to the TEM grid and washed three times with water. After washing, the TEM grid was stained with 1% aq. Phosphotungstic acid, PTA (negative stain), then air dried and observed under TEM for imaging. The image was captured with a Jeol 1400 Plus TEM at 120 kV and 15000X magnification. The positive cell suspension was found positive for FMD virus under TEM in which virion capsid appears to be icosahedral, whereas in mock infected cell suspension samples no FMD virus particles were observed. The current study describes the process of demonstration of FMD virion in cell suspension using a negative staining method. This method can also be used to detect FMD virus in vesicular fluid, other fluids suspected for FMD virus, and other viruses.

Keywords: Cell Suspension; Foot-and-Mouth Disease Virus; Negative Staining; Phosphotungstic Acid; Transmission Electron Microscope

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Citation

Citation: Rajeev Ranjan., et al. “Optimization and Development of Protocol for Detection of Foot-And-Mouth Disease Virus by Negative Staining Using Transmission Electron Microscopy".Acta Scientific Veterinary Sciences 5.2 (2023): 18-23.

Copyright

Copyright: © 2023 Rajeev Ranjan., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.




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