Rare occurrence of Coxsackievirus B4 along with Coxsackieviruses A6 and A10 from hand, foot and mouth disease

Background: Hand, Foot and Mouth Disease (HFMD) has been widely spread in Asia and resulted in a high disease burden among children in many countries. It remains a common problem in India, yet its etiology is largely unknown as diagnosis is based on clinical characteristics. Aim and Objectives: To investigate the circulation of strains, clinical, molecular and virological aspects of the disease. Material and Methods: Vesicular lesion and throat swab specimens were referred from eight children for laboratory diagnosis of HFMD. The detection of enteroviruses was performed by nested PCR, virus isolation using HeLa cell line and sequencing of 5 ' UTR. Results: Coxsackieviruses A6, A10 and B4 were identified by nested PCR, isolated in HeLa cell line and reconfirmed. The phylogenetic analysis of Coxsackieviruses A6, A10 and B4 showed similarity with Beijing-China, Pune-India, and France respectively. Conclusion: The findings of this study indicate the co-circulation of Coxsackieviruses A6, A10 and B4. Coxsackievirus B4 was isolated and detected for the first time from Pune, India. Additionally, results emphasize the importance of virus isolation using cell lines for laboratory diagnosis. Point-of-Care testing and vaccine development would be beneficial towards control and prevention of HFMD.