Comparison of Classic and Molecular Approaches for the Identification of Untypeable Enteroviruses
| Autor: | Mark A. Pallansch, G Marchetti, David R. Kilpatrick, Mary R. Flemister, M S Oberste, Kaija Maher |
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| Rok vydání: | 2000 |
| Předmět: |
Microbiology (medical)
Serotype Picornavirus viruses Molecular Sequence Data ved/biology.organism_classification_rank.species Enterovirus C Picornaviridae medicine.disease_cause Sensitivity and Specificity law.invention Mice Antibody Specificity Neutralization Tests law Virology Enterovirus Infections Antigenic variation medicine Animals Humans Horses Typing Serotyping Polymerase chain reaction Enterovirus Mammals Picornaviridae Infections biology Reverse Transcriptase Polymerase Chain Reaction ved/biology Immune Sera virus diseases biology.organism_classification Rabbits Rhinovirus |
| Zdroj: | Journal of Clinical Microbiology. 38:1170-1174 |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.38.3.1170-1174.2000 |
| Popis: | Members of the family Picornaviridae are the most common viruses infecting humans, and species in several genera also infect a wide variety of other mammals. Picornaviruses have traditionally been classified by antigenic type, based on a serum neutralization assay. However, this method is time-consuming and labor-intensive, is sensitive to virus aggregation and antigenic variation, and requires a large number of antisera to identify all serotypes, even when antiserum pools are used. We developed generic reverse transcription (RT)-PCR primers that will amplify all human enterovirus serotypes, as well as many rhinoviruses and other picornaviruses, and used RT-PCR amplification of the VP1 gene and amplicon sequencing to identify enteroviruses that were refractory to typing by neutralization with pooled antisera. Enterovirus serotypes determined by sequencing were confirmed by neutralization with monospecific antisera. Of 55 isolates tested, 49 were of known enterovirus serotypes, two were rhinoviruses, and four were clearly picornaviruses but did not match any known picornavirus sequence. All four untyped picornaviruses were closely related to one another in sequence, suggesting that they are of the same serotype. RT-PCR, coupled with amplicon sequencing, is a simple and rapid method for the typing and classification of picornaviruses and may lead to the identification of many new picornavirus serotypes. |
| Databáze: | OpenAIRE |
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