ASSOCIATION OF NSP4 MUTATION AND VP4 ROTAVIRUS GENOTYPE IN CHILDREN WITH SEVERE-MODERATE DIARRHEA
Elsa Fitri Ana, Rury Mega Wahyuni, Laura Navika Yamani, Zayyin Dinana, Devi Oktafiani, Ni Luh Ayu Megasari, Maria Inge Lusida, Juniastuti, Soetjipto

The Doctoral Program, Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia-

2 Institute of Tropical Disease, Universitas Airlangga, Surabaya, Indonesia-

3 Department of Medical Biochemistry, Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia-

4 Department of Microbiology, Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia-

5 Indonesia-Japan Collaborative Research Center for Emerging and Re-emerging Infectious Diseases, Universitas Airlangga, Surabaya, Indonesia-

6Community Education, Faculty of Education, Universitas Negeri Jakarta, Jakarta, Indonesia

Abstract

Rotavirus is the main cause of severe acute diarrhea among children. NSP4 has a role in the development of secretory diarrhea. NSP4 genotype is divided into 21 genogroups (E1-E21). Information related to rotavirus infection and the role of NSP4 pathogenesis in human has not been described as clear information about the association between NSP4 and VP4 is still limited. To determine the genotype and mutation of NSP4 and to analyze the mutation of NSP4 with VP4 genotype and diarrhea severity. A total of 51 rotavirus-positive samples were collected from children with acute diarrhea between August-Dec 2016 and June-August 2018 from 2 hospitals and 2 primary healthcare in Bandar Lampung, Indonesia. The severity of diarrhea data was collected by observation, interviewing parents or guardians, and from medical records based on Ruuska and Vesikari score. Identification of VP4 gene was performed using primers from the WHO Manual of RV detection and characterization methods protocols. P types of these samples were determined by sequencing methods. Amplification DNA for NSP4 gene was carried out using the primers described before. Amplified viral genes were subjected to sequence analysis. In this study, amino acids substitution was found in H1, H2, H3 domain, enterotoxin region, VP4-binding site, and VP6-binding site from E1 and E2 genotype. Unusual combination occured between P[8] genotype and E2 with amino acid substitution at position 72 in children with severe diarrhea. The enterotoxin region mutation at position 129 and hydrophobic domain H1 at position 14 mostly occurred in NSP4 sequence associated with P[4] and in children with severe diarrhea. This finding indicates that amino acids variation in NSP4 sequence related to VP4 genotype, but no association with diarrhea severity.

Keywords: NSP4, mutation, VP4 genotype, severity

Topic: Medical : basic science, clinical, translational research, medical education, and miscellaneous