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RV011 Subtyping and investigation of transmission events of Rotavirus type A using phylogenetic analysis
Wissenschaftliches Programm 55. DGHM-Tagung 29. September-1. Oktober 2003 in Dresden Abstracts - Kurzvortr~ge than 0.5 % and finally led to the establishment of national and European recommendations on the guidance and management of health-care workers infected with HCV. (1) Clin Lab Anal 2001; 1 5 : 3 0 8 - 3 1 3 (2) J Virol Methods 2002; 101:159-168 (3) J Clin Microbiol 2000; 38:3581-3584 (4) J Med Virol 2003; 69:331-338 (5) NEJM 2000; 343:1851-1854 (6) Arch Intern Med 2002; 162:805-810 (7) J Med Virol 2002; 66:461-467
Subtyping and investigation of transmission events of Rotavirus Type A using phylogenetic analysis St0rmer, M. 1, Unverzagt, H.1; Rabenau, H.1; Doerr, H.W} ~Universit~tsklinikum Frankfurt/Nain ; InsUtut fDr Medizinische Virologie
Rotavirus infection is associated with human gastroenteritis, and is the major cause of infantile gastroenteritis worldwide. In the hospital environment, Rotavirus infections often occur nosocomially. Outbreaks were mainly analysed using serological or PCR subtyping methods (VP7 and VP4 gene). Here we describe a phylogenetic analysis including 28 VP4 and 29 VP7 GenBank sequences using a single PCR product to determine Rotavirus A subtypes, and are planning to investigate possible nosocomial outbreaks. Rotavirus RNA was extracted using the QIAamp Viral RNA Mini Kit (Qiagen, Hilden, Germany). RT-PCR was performed using the OneStep RT-PCR Kit (Qiagen) with primers Con2/Con3 (VP4; Gentsch, 1992), and Beg9/End9 (VP7; Gouvea, 1990). PCR products were sequenced with specific primers, and analysed on an ABI 377-96 sequencer (Applied Biosystems, Darmstadt, Germany). Sequence alignments were done using the MegAlign module of the LaserGene software (DNASTAR, Madison, WI, USA), and imported into the Mega2 software (http ://www.megasoftware. net) for phylogenetic analysis. The different sero- and genogroups of the GenBank isolates were clearly distinguishable in the phylogenetic analysis for both the VP4 and VP7 gene. Patient isolates could definitely be assigned to a specific geno- or serotype defined by a subtype-specific cluster of the GenBank isolates. The patient isolates had no common source of infection, as they were clearly unrelated in the phylogenetic trees. Our results show that subtyping of Rotavirus sero- and genotypes can be performed very easily using phylogenetic analysis, and that investigations of nosocomial outbreaks can be done in parallel to the subtyping procedure.
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Subtyping and investigation of transmission events of Rotavirus Type A using phylogenetic analysis St0rmer, M. 1, Unverzagt, H.1; Rabenau, H.1; Doerr, H.W} ~Universit~tsklinikum Frankfurt/Nain ; InsUtut fDr Medizinische Virologie
Rotavirus infection is associated with human gastroenteritis, and is the major cause of infantile gastroenteritis worldwide. In the hospital environment, Rotavirus infections often occur nosocomially. Outbreaks were mainly analysed using serological or PCR subtyping methods (VP7 and VP4 gene). Here we describe a phylogenetic analysis including 28 VP4 and 29 VP7 GenBank sequences using a single PCR product to determine Rotavirus A subtypes, and are planning to investigate possible nosocomial outbreaks. Rotavirus RNA was extracted using the QIAamp Viral RNA Mini Kit (Qiagen, Hilden, Germany). RT-PCR was performed using the OneStep RT-PCR Kit (Qiagen) with primers Con2/Con3 (VP4; Gentsch, 1992), and Beg9/End9 (VP7; Gouvea, 1990). PCR products were sequenced with specific primers, and analysed on an ABI 377-96 sequencer (Applied Biosystems, Darmstadt, Germany). Sequence alignments were done using the MegAlign module of the LaserGene software (DNASTAR, Madison, WI, USA), and imported into the Mega2 software (http ://www.megasoftware. net) for phylogenetic analysis. The different sero- and genogroups of the GenBank isolates were clearly distinguishable in the phylogenetic analysis for both the VP4 and VP7 gene. Patient isolates could definitely be assigned to a specific geno- or serotype defined by a subtype-specific cluster of the GenBank isolates. The patient isolates had no common source of infection, as they were clearly unrelated in the phylogenetic trees. Our results show that subtyping of Rotavirus sero- and genotypes can be performed very easily using phylogenetic analysis, and that investigations of nosocomial outbreaks can be done in parallel to the subtyping procedure.
http:llwww.dghm,org
196