Adaptation of Three Different BLRI Strain (O, A, Asia 1) of Foot and Mouth Disease Virus Serotypes in Baby Hamster Kidney-21 Cell Line

Mohammad Showkat Mahmud,Md Rezaul Karim,Eusha Islam,Md Giasuddin,Amal Kumar Saha,Mohammed Abdus Samad

doi:10.13189/iid.2017.050201

Abstract

Baby Hamster Kidney-21(BHK-21) cells are the most commonly used cell for viral seed preparation and vaccine production. The aim of the present study was to adapt BLRI three different field isolates of foot and mouth disease virus (FMDV) serotypes (O, A, Asia 1) into baby hamster kidney-21 cell line for FMD trivalent master seed preparation. For this, samples were collected from the tongue epithelium, saliva, milk and inter digital tissues of the suspected animals from different districts of Bangladesh during the year 2011-2014. All samples were subjected to RNA extraction and RT-PCR for detection and serotyping of FMDV. BHK-21 cell subculture was done from a previously cultured flask containing BHK-21 cells. Prepared inocula (positive for serotype O, A, Asia 1) were inoculated into BHK-21 cell culture and cytopathic effects (CPE) were observed in BHK-21 cell line characterized by rounding and flattening of the cells, multinucleated giant cells formation, breaking down of the intracellular bridges and finally cell death indicated the presence of FMDV. Adaptation of the virus into baby hamster kidney-21 cell line was confirmed by detection of viral infection in infected cells by RT-PCR with serotype specific primers. The cell adapted foot and mouth disease virus in the present study provides a scope to undertake further research on foot and mouth disease virus in Bangladesh towards development of trivalent FMD vaccine for use in the country.

Highlights

Foot and mouth disease (FMD) is considered a highly infectious disease of cattle, sheep, goats, and pigs
It was found that 73.13% samples were positive for FMD virus out of 134 suspected samples, 31 samples were found positive for foot and mouth disease virus (FMDV) serotype O, 7 were positive for FMDV serotype A, 30 were positive for FMDV serotype Asia 1, 9 samples were positive for mixed infection and 21 were nontype following Reverse Transcription Polymerase Chain reaction (RT-PCR) using serotype specific primer (Figure 1)
Cytopathic effects (CPE) were observed in baby hamster kidney (BHK)-21 cell line characterized by rounding and flattening of the cells, multinucleated giant cells formation, breaking down of the intracellular bridges and cell death indicated the presence of FMDV (Figure 3)

Summary

Introduction

Foot and mouth disease (FMD) is considered a highly infectious disease of cattle, sheep, goats, and pigs. It affects wild animals such as buffaloes and deer [1, 2].

Materials and Methods

Sample Collection

Subculture of BHK-21 Cell

Adaptation of FMDV in BHK-21 Cell Line

Results and Discussion

Conclusions