Bharath Mudurangaplar1, B V Peerapur2. 1. PhD Scholar, Department of Microbiology, Shri B.M. Patil Medical College and Research Center, BLDE University , Bijapur, Karnataka, India . 2. Professor and HOD, Department of Microbiology, Raichur Institute of Medical Sciences (RIMS) , Raichur, Karnataka, India .
Abstract
INTRODUCTION: Indian ocean islands and India have experienced massive severe Chikungunya outbreak from 2005 up till now and then Chikungunya became epidemic in India. The mutations that occurred in E1 gene were responsible for increased infectivity, virulence and host adaptability. It is important to find out the genotype and its probable evolvement and novel mutations in the E1 gene reported during 2006-2009 from the current isolates, which may affect the local protein structure. AIM: To perform Molecular diagnosis and Molecular Characterisation of Chikungunya virus isolates. MATERIALS AND METHODS: A total of 33 samples were included in the study. RNA was isolated from 33 serum samples and Real time PCR was carried out. Further, Nested PCR and E1 partial gene sequencing was performed. Phylogenetic analysis, mutational analysis and protein modelling studies were carried out. RESULTS: Out of 33 samples tested, 31 were found positive for CHIK RNA. Phylogenetic analysis showed that isolates belongs to ECSA genotype and E1K211E, E1M269V and E1D284E mutations were observed from all the isolates. CONCLUSION: The isolates may have evolved from ECSA Reunion island strains and identified unique mutations in E1 gene were maintained. These mutations have not affected local protein structure.
INTRODUCTION: Indian ocean islands and India have experienced massive severe Chikungunya outbreak from 2005 up till now and then Chikungunya became epidemic in India. The mutations that occurred in E1 gene were responsible for increased infectivity, virulence and host adaptability. It is important to find out the genotype and its probable evolvement and novel mutations in the E1 gene reported during 2006-2009 from the current isolates, which may affect the local protein structure. AIM: To perform Molecular diagnosis and Molecular Characterisation of Chikungunya virus isolates. MATERIALS AND METHODS: A total of 33 samples were included in the study. RNA was isolated from 33 serum samples and Real time PCR was carried out. Further, Nested PCR and E1 partial gene sequencing was performed. Phylogenetic analysis, mutational analysis and protein modelling studies were carried out. RESULTS: Out of 33 samples tested, 31 were found positive for CHIK RNA. Phylogenetic analysis showed that isolates belongs to ECSA genotype and E1K211E, E1M269V and E1D284E mutations were observed from all the isolates. CONCLUSION: The isolates may have evolved from ECSA Reunion island strains and identified unique mutations in E1 gene were maintained. These mutations have not affected local protein structure.
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