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EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR
Published on: January 16, 2016
Molecular identification and characterization of two proposed new enterovirus serotypes, EV74 and EV75
M Steven Oberste1, Suzanne M Michele1, Kaija Maher1
1Respiratory and Enteric Viruses Branch, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road NE, Mailstop G-17, Atlanta, GA 30333, USA.
Abstract:
Sequencing of the gene that encodes the capsid protein VP1 has been used as a surrogate for antigenic typing in order to distinguish enterovirus serotypes; three new serotypes were identified recently by this method. In this study, 14 enterovirus isolates from six countries were characterized as members of two new types within the species Human enterovirus B, based on sequencing of the complete capsid-encoding (P1) region. Isolates within each of these two types differed significantly from one another and from all other known enterovirus serotypes on the basis of sequences that encode either VP1 alone or the entire P1 region. Members of each type were > or =77.2 % identical to one another (89.5 % amino acid identity) in VP1, but members of the two different types differed from one another and from other enteroviruses by > or =31 % in nucleotide sequence (25 % amino acid sequence difference), indicating that the two groups represent separate new candidate enterovirus types. The complete P1 sequences differed from those of all other enterovirus serotypes by > or =31 % (26 % amino acid sequence difference), but were highly conserved within a serotype (<8 % amino acid sequence difference). Phylogenetic analyses demonstrated that isolates of the same serotype were monophyletic in both VP1 and the capsid as a whole, as shown previously for other enterovirus serotypes. This paper proposes that these 14 isolates should be classified as members of two new human enterovirus types, enteroviruses 74 and 75 (EV74 and EV75).
Insights
Two new human enterovirus types, enteroviruses 74 and 75 (EV74 and EV75), were identified using capsid protein VP1 gene sequencing. These findings expand the classification of human enteroviruses within the Human enterovirus B species.
Area of Science:
- Virology
- Molecular Biology
- Public Health
Background:
- Enterovirus serotypes are traditionally distinguished by antigenic typing.
- Sequencing of the capsid protein VP1 gene has emerged as a reliable method for enterovirus identification.
- Previous studies have identified new enterovirus serotypes using VP1 sequencing.
Purpose of the Study:
- To characterize 14 enterovirus isolates from six countries.
- To determine if these isolates represent new enterovirus types within the Human enterovirus B species.
- To propose a classification for these novel enterovirus types.
Main Methods:
- Sequencing of the complete capsid-encoding (P1) region of 14 enterovirus isolates.
- Comparative sequence analysis of the P1 region and the VP1 gene.
- Phylogenetic analysis to determine evolutionary relationships.
Main Results:
- Two distinct groups of enterovirus isolates were identified, representing two new candidate types.
- These new types, proposed as enteroviruses 74 and 75 (EV74 and EV75), showed significant sequence divergence from known enteroviruses.
- Sequence analysis of the P1 region and VP1 gene confirmed high conservation within each type and significant differences between types.
- Phylogenetic analyses confirmed the monophyletic nature of isolates within each proposed serotype.
Conclusions:
- The 14 characterized isolates represent two novel human enterovirus types, EV74 and EV75.
- VP1 and P1 region sequencing are effective methods for identifying and classifying new enterovirus types.
- These findings contribute to the ongoing classification and understanding of the enterovirus genus.
