Rapid molecular epidemiologic studies of human parainfluenza viruses based on direct sequencing of amplified DNA from a multiplex RT-PCR assay
| Autor: | Dean D. Erdman, Larry J. Anderson, J E Echevarría, H C Meissner |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Paramyxoviridae
Sequence analysis Molecular Sequence Data Virus law.invention law Virology Humans Multiplex Child Polymerase chain reaction Genetics Molecular Epidemiology HN Protein Paramyxoviridae Infections biology Reverse Transcriptase Polymerase Chain Reaction Sequence Analysis DNA biology.organism_classification Reverse transcriptase Parainfluenza Virus 1 Human Parainfluenza Virus 2 Human Parainfluenza Virus 3 Human Reverse transcription polymerase chain reaction Real-time polymerase chain reaction DNA Viral |
| Zdroj: | Journal of virological methods. 88(1) |
| ISSN: | 0166-0934 |
| Popis: | Sequencing studies of limited regions of the human parainfluenza viruses (HPIVs) genomes have helped describe patterns of virus circulation and characterize institutional outbreaks of HPIVs-associated respiratory illness. In this study, we sequenced reverse transcription polymerase chain reaction (RT-PCR)-amplified HPIVs RNA obtained from a multiplex RT-PCR assay described previously for simultaneous detection of HPIV-1, 2 and 3. Differences in the nucleotide sequences of limited regions of the HN gene allowed us to distinguish temporally and geographically diverse HPIV isolates (43 HPIV-1, 7 HPIV-2, 12 HPIV-3 isolates from this and previously published studies). In addition, an outbreak of HPIV-3-associated illness among infants on a pediatric ward was investigated by comparing sequences of three ward isolates with three matched community controls. Sequences of all ward isolates were identical and differed from those of the community controls, suggesting a single introduction and nosocomial transmission of the virus. Combining multiplex reverse transcription polymerase chain reaction (RT-PCR) assays with direct sequencing of the PCR products can provide an integrated system for rapid diagnosis and characterization of HPIVs. |
| Databáze: | OpenAIRE |
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