Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Isolation and molecular characterization of coxsackievirus A6 and coxsackievirus A16 from a case of recurrent Hand, Foot and Mouth Disease in Mumbai, Maharashtra, India, 2018

  • 10 Accesses


Hand, Foot and Mouth Disease (HFMD) is caused by multiple Enterovirus (EV) serotypes mainly coxsackievirus A6 (CV-A6), coxsackievirus A16 (CV-A16) and Enterovirus 71 (EV-A71). Recurrent HFMD infections are rarely reported. An unusual rise in HFMD cases was reported in Mumbai during May–June 2018. Stool and throat swab specimens were referred from seven children from two hospitals for laboratory diagnosis. The age group of cases ranged from 9 months to 5 years with median age 13 months. Out of seven cases, three were males and four females. One 13-month-old female case was reported twice within 21 days. Stool, throat swab specimens were tested by pan enterovirus RT-PCR and also by virus isolation using human rhabdomyosarcoma cell line for detection of Enteroviruses. Out of seven HFMD cases, CV-A6 and CV-A16 viruses were isolated from five and two cases respectively. The phylogenetic analysis of CV-A6 viruses showed their similarity with CV-A6 viruses from Finland and China, whereas the two CV-A16 isolates showed similarity with those from Japan, France, China, Sarawak and Thailand. For the recurrent HFMD case, CV-A6 and CV-A16 were isolated from the stool specimens collected during the first and second episodes, respectively. There are no reports of isolation and molecular characterization of CV-A6 and CV-A16 viruses from recurrent HFMD cases. The present study reports molecular characterization of two Enterovirus serotypes CV-A6 and CV-A16 from a recurrent HFMD case, highlighting need of virological and molecular surveillance of HFMD.

This is a preview of subscription content, log in to check access.

Fig. 1


  1. 1.

    Ang LW, Koh BK, Chan KP, Chua LT, James L, Goh KT. Epidemiology and control of hand, foot and mouth disease in Singapore, 2001–2007. Ann Acad Med Singapore. 2009;38:106–12.

  2. 2.

    Dharmapalan D, Saxena VK, Pawar SD, Qureshi THIH, Surve P. Clinical and molecular investigations of hand, foot and mouth disease outbreak in Navi Mumbai, India. Indian Pediatr. 2019;56:1052–4.

  3. 3.

    Dias EDM, Dias M. Recurring hand foot mouth disease in a child. Ann Trop Med Public Health. 2012;5:40–1. https://doi.org/10.4103/1755-6783.92879.

  4. 4.

    Gopalkrishna V, Patil PR, Patil GP, Chitambar SD. Circulation of multiple enterovirus serotypes causing hand, foot and mouth disease in India. J Med Microbiol. 2012;61:420–5. https://doi.org/10.1099/jmm.0.036400-0.

  5. 5.

    Hosoya M, Kawasaki Y, Sato M, Honzumi K, Hayashi A, Hiroshima T, et al. Genetic diversity of coxsackievirus A16 associated with hand, foot, and mouth disease epidemics in Japan from 1983 to 2003. J Clin Microbiol. 2007;45:112–20. https://doi.org/10.1128/JCM.00718-06.

  6. 6.

    Huang WC, Huang LM, Kao CL, Lu CY, Shao PL, Cheng AL, et al. Seroprevalence of enterovirus 71 and no evidence of cross-protection of enterovirus 71 antibody against the other enteroviruses in kindergarten children in Taipei city. J Microbiol Immunol Infect. 2012;45:96–101. https://doi.org/10.1016/j.jmii.2011.09.025.

  7. 7.

    Huang J, Liao Q, Ooi MH, Cowling BJ, Chang Z, Wu P, et al. Epidemiology of recurrent hand, foot and mouth disease, China, 2008–2015. Emerg Infect Dis. 2018;24:432–42. https://doi.org/10.3201/eid2403.171303.

  8. 8.

    Kar BR, Dwibedi B, Kar SK. An outbreak of hand, foot and mouth disease in Bhubaneswar, Odisha. Indian Pediatr. 2013;50:139–42.

  9. 9.

    Miller GD, Tindall JP. Hand-foot-and-mouth disease. JAMA. 1968;203:827–30. https://doi.org/10.1001/jama.1968.03140100009002.

  10. 10.

    Nix WA, Oberste MS, Pallansch MS. Sensitive, Semi-nested PCR amplification of VP1 sequences for direct identification of all Enterovirus serotypes from original Clinical specimens. J Clin Microbiol. 2006;44:2698–704. https://doi.org/10.1128/JCM.00542-06.

  11. 11.

    Oberste MS, Maher K, Flemister MR, Marchetti G, Kilpatrick DR, Pallanch MA. Comparison of classic and molecular approaches for the identification of untypeable enteroviruses. J Clin Microbiol. 2000;38:1170–4.

  12. 12.

    Oberste MS, Maher K, Kilpatrick DR, Pallansch MA. Molecular evolution of the human enteroviruses: correlation of serotype with VP1 sequence and application to picornavirus classification. J Virol. 1999;73:1941–8.

  13. 13.

    Oberste MS, Nix WA, Maher K, Pallanch M. Improved molecular identification of enteroviruses by RT-PCR and amplicon sequencing. J Clin Microbiol. 2003;26:375–7. https://doi.org/10.1016/S1386-6532(03)00004-0.

  14. 14.

    Sabhita S, Sasidharanpillai S, Sanjay RE, Binitha MP, Riyaz N, Muhammed K, et al. Clinical profile and virology analysis of hand, foot and mouth disease from North Kerala, India in 2015–2016: a tertiary care hospital based cross-sectional study. Indian J Dermatol Venereol Leprol. 2018;84:328–31. https://doi.org/10.4103/ijdvl.IJDVL_579_17.

  15. 15.

    Sane S, Saboo N, Utange V. Observations regarding outbreak of hand, foot and mouth disease at Thane, Maharashtra. Pediatr Infect Dis. 2009;1:110–1. https://doi.org/10.1016/S2212-8328(09)80013-1.

  16. 16.

    Saoji VK. Hand, foot and mouth disease in Nagpur. Indian J Dermatol Venereol Leprol. 2008;74:133–5. https://doi.org/10.4103/0378-6323.39697.

  17. 17.

    Sarma N, Chakraborty S, Dutta A, Sadhukhan PC. Hand, foot and mouth disease in West Bengal, India: a preliminary report on clinico virological trend over 3 successive years (2013–2015). Indian J Dermatol. 2017;62:486–90. https://doi.org/10.4103/ijd.IJD_381_17.

  18. 18.

    Sasidharan CK, Sugathan P, Agarwal P, Khare SL, Jayaram Paniker CK. Hand-foot-and-mouth disease in Calicut. Ind J Pediatr. 2005;72:17–21. https://doi.org/10.1007/BF02760573.

  19. 19.

    Sutton-Hayes S, Weisse ME, Wilson NW, Ogershok PR. A recurrent presentation of hand, foot, and mouth disease. Clin Pediatr. 2006;45:373–6. https://doi.org/10.1177/000992280604500412.

  20. 20.

    World Health Organization. Polio laboratory manual. 4th ed. Geneva: World Health Organization; 2004.

  21. 21.

    Yadav PD, Albarino CG, Nyayanit DA, Guerrero L, Jenks MH, Sarkale P, et al. Equine Encephalosis virus in India, 2008. Emerg Infect Dis. 2018;24:898–901. https://doi.org/10.3201/eid2405.171844.

  22. 22.

    Yang CF, De L, Yang SJ, Gomez JR, Cruz JR, Holloway BP, et al. Genotype-specific in vitro amplification of sequences of the wild type 3 polioviruses from Mexico and Guatemala. Virus Res. 1992;24:277–96. https://doi.org/10.1016/0168-1702(92)90124-R.

  23. 23.

    Zhu FC, Meng FY, Li JX, Li XL, Mao QY, Tao H, et al. Efficacy, safety, and immunology of an inactivated alum-adjuvant enterovirus 71 vaccine in children in China: a multicentre, randomised, double-blind, placebo-controlled, phase 3 trial. Lancet. 2013;381:2024–32. https://doi.org/10.1016/S0140-6736(13)61049-1.

Download references


We sincerely thank Dr. DT Mourya, Former Director, NIV, Pune for his keen interest in the work and support. We also thank Dr. RR Gangakhedkar, Head, ECD, Dr. Nivedita Gupta, Scientist-F, ECD, Indian Council of Medical Research Head Quarters, New Delhi for prompting this study.


This work was supported by intramural grant from the Indian Council of Medical Research.

Author information

Correspondence to Vinay K. Saxena or Shailesh D. Pawar.

Ethics declarations

Conflict of interest

The author(s) declare that they have no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Saxena, V.K., Pawar, S.D., Qureshi, T.H.I.H. et al. Isolation and molecular characterization of coxsackievirus A6 and coxsackievirus A16 from a case of recurrent Hand, Foot and Mouth Disease in Mumbai, Maharashtra, India, 2018. VirusDis. (2020). https://doi.org/10.1007/s13337-020-00567-1

Download citation


  • Hand Foot and Mouth Disease
  • Recurrence
  • Coxsackievirus
  • Enteroviruses