Rotavirus Gastroenteritis and Central Nervous System (CNS) Infection: Characterization of the VP7 and VP4 Genes of Rotavirus Strains Isolated from Paired Fecal and Cerebrospinal Fluid Samples from a Child with CNS Disease

Abstract

Rotavirus RNA was detected in the cerebrospinal fluid (CSF) of a child with central nervous system disease symptoms associated with rotavirus gastroenteritis. The rotavirus isolates from the fecal and CSF samples were genotyped as G1P [8]. Sequence analysis of the VP7 and VP4 proteins derived from the fecal and CSF samples were remarkably similar to each other and to G1P[8] rotavirus strains commonly circulating in the community and associated with gastroenteritis. CASE REPORTA 22-month-old previously healthy male child was admitted to the hospital with a 2-day history of nonbloody diarrhea and vomiting. On the day of admission he developed fever and had a generalized tonic-clonic seizure lasting 2 min. On arrival at the hospital, he was apyrexial, mildly dehydrated, and drowsy. Physical examination was otherwise unremarkable with no neck stiffness.A lumbar puncture was performed. The cerebrospinal fluid (CSF) contained 1,235 red cells/l and eight white cells/l (75% lymphocytes). A blood count showed a total white cell count of 9.4 ϫ 10 9 cells/liter, with 2.4 ϫ 10 9 lymphocytes/liter and 6.0 ϫ 10 9 neutrophils/liter (normal for the patient's age). Intravenous penicillin, cefotaxime, and acyclovir were administered. Bacterial cultures of blood and CSF were negative, and antibiotic treatment was stopped after 72 h. Clinically the child improved over the first 24 h and subsequently made a full recovery.Microbiologic analysis. A stool sample was positive for rotavirus antigen by latex particle agglutination, and this result was confirmed by inhibition with specific antibodies. Both stool and CSF samples were tested for the presence of rotavirus RNA. Nucleic acid was extracted and reverse transcribed, and the cDNA was used in rotavirus VP7-and VP4-specific typing PCRs as described previously (7). Both the stool and CSF samples contained rotavirus of the genotype G1P [8]. The VP7 and VP4 first-round amplicons derived from the stool and CSF samples were purified and sequenced in both directions by using the same consensus primers as those used for the firstround amplification. Nucleic acid sequences were derived using an automated sequencer (CEQ2000; BeckmanCoulter). Sequence data were analyzed using the software programs Seqman and Megalign (DNA Star; Lasergene, Madison, Wis.).Partial cDNA sequences of the genes encoding the rotavirus VP7 (nucleotides [nt] 51 to 932) and VP4 (nt 11 to 887) obtained from the stool and CSF samples were highly homologous (Ͼ98.5 and 100% at the nucleotide and amino acid levels, respectively). The sequences of VP7 and VP4 clustered within the G1 genetic lineage III and the P[8] genetic lineage II, respectively ( Fig. 1 and 2) (8, 9). Analysis of the deduced amino acid sequences revealed no amino acid substitutions between the VP7 amino acid sequences derived from the fecal and CSF samples and VP7 sequences of other rotavirus strains within their genetic lineage. The deduced amino acid sequence of VP4 revealed a single amino acid substitution at position 166 within the hypervariable region...

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