Background: Rota vaccine is used to prevent diarrhea in children under 5 years old. Two vaccines are being used in developed countries: Rotarix (GSK) and RotaTeq (Merk). Rotarix vaccine was produced from master seed G1P8 and RotaTeq vaccine was from the coordination of human rotavirus strains G1, G2, G3, G4 and cow rotavirus strain. Thanks to the help of WHO, Ministry of Health and Ministry of Science and Technology, Center for research and production of vaccines and biologicals \ufffd?Ha Noi made study of creating rotavirus master seed G4P6 for Rota vaccine production in Vietnam. Objectives: To evaluate the safety and potency of rotavirus master seed G4P6 in the laboratory and experimental animals. Subjects and method: Rotavirus master seed G4P6 (2001019203) lot 1 (MS-PL5) and lot 2 (MS-PL5) produced in 2005, preserved at -800C were determined potency by Immunofluorescence (IF) method and tested for safety on rabits and rats. Results:2 lots of Rotavirus master seed G4P6 that had been produced in Center for research and production of vaccines and biologicals \ufffd?Ha Noi had high titre and safety in the laboratory and experimental animals. Conclusion: The results were the basis of Rota vaccine production in Vietnam.
Rotavirus/ isolation & ; purification ; Rotavirus Vaccines/ isolation & ; purification ; contraindications

Background: Rota vaccine is used to prevent diarrhea in children under 5 years old. Two vaccines are being used in developed countries: Rotarix (GSK) and RotaTeq (Merk). Rotarix vaccine was produced from master seed G1P8 and RotaTeq vaccine was from the coordination of human rotavirus strains G1, G2, G3, G4 and cow rotavirus strain.Thanks to the helps of WHO, Ministry of Health and Ministry of Science and Technology, Center for research and production of vaccines and biologicals \ufffd?Ha Noi made study of creating rotavirus master seed G1P4 for Rota vaccine production in Vietnam. Objectives: To evaluate the safety and potency of rotavirus master seed G1P4 in the laboratory and experimental animals. Subjects and method: Rotavirus master seed G1P4 (2001019210) lot 1 (MS-P5) and lot 2 (MS-P5) produced in 2005, preserved at -800C were determined potency by Immunofluorescence (IF) method and tested for safety on rabits and rats. Results:2 lots of Rotavirus master seed G1P4 that had been produced in Center for research and production of vaccines and biologicals \ufffd?Ha Noi had high titre and safety in the laboratory and experimental animals. Conclusion: The result was the basis of Rota vaccine production in Vietnam.
Rotavirus/ diagnostic use ; Rotavirus Vaccines/ isolation & ; purification ; contraindications

China ; epidemiology ; Genes, Viral ; Humans ; Polymerase Chain Reaction ; methods ; Rotavirus ; genetics ; isolation & purification ; Rotavirus Infections ; epidemiology ; virology

Rotavirus is the leading causal agent of severe acute gastroenteritis in children aged <5 years. A specific pharmacologic agent for the treatment of rotavirus-infected children is lacking. In China, only the Luo Tewei oral vaccine (Lanzhou Institute of Biological Products, Shanghai, China), which is produced from Lanzhou lamb rotavirus vaccine (LLR), is available. Studies have hypothesized that the genotype of LLR is G10P[12], To identify the genotype of LLR by reverse transcription-polymerase chain reaction, we showed that the VP7 and VP4 genotypes of LLR were G10 and P[15], respectively, based on sequencing, alignment and phylogenetic analyses. In conclusion, we identified the genotype of rotavirus strain LLR to be G10P[15].
China ; Genotype ; Humans ; Molecular Sequence Data ; Phylogeny ; Rotavirus ; chemistry ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; virology ; Rotavirus Vaccines ; chemistry ; classification ; genetics ; isolation & purification ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics

To develop a specific, rapid and convenient method based on molecular motor biosensor to detect food-borne rotavirus. A specific probe was encompassed the conservative region of rotavirus's VP7 segment, and a molecular motor detect device was constructed by connecting probes to F0F1-ATPase molecular motor through biotin-streptavidin system. This biosensor's sensitivity was 0.005 ng/mL for rotavirus RNA. Extracted virus RNA was conjugated with the biosensor separately, at the same time ATP was synthesized. By comparing fluorescence intensity, we can detect rotavirus RNA in samples. This method possessed specificity for rotavirus, without any cross-reaction with Hepatitis A virus and noroviris, and it could be accomplished within 1 h. We detected 15 samples using this method and the results were compared with RT-PCR results. This method is sensitive and specific for rotavirus, and it can be used to detect food-borne rotavirus.
Biosensing Techniques ; methods ; DNA, Viral ; analysis ; genetics ; Food Microbiology ; methods ; Rotavirus ; genetics ; isolation & purification ; Sensitivity and Specificity

Child ; Child, Preschool ; Diarrhea ; virology ; Female ; Genotype ; Humans ; Infant ; Male ; Rotavirus ; classification ; genetics ; isolation & purification ; Seasons

P[8]b is a newly discovered sub-genotype for VP4 gene of group A human rotaviruses (HRV) worldwide. This study was to develop an effective method to identify P[8]a, P[8]b, P[4] and P[6] (sub) genotypes of VP4 genes of HRV and to investigate the prevalence of P[8]b sub-genotype and its G/P combinations of HRV in outpatient and inpatient children with diarrhea in Children's Hospital affiliated to Capital Institute of Pediatrics from 2009 to 2010. By analyzing the collected nucleotide sequences of VP4 gene for all known P genotypes of HRV including P[8]b subtype from GenBank and using softwares of DNAS-tar and MegAlign to align and analyze multiple sequences, probes for P[8]a, P[8]b, P[4] and P[6] (sub) genotypes in the corresponding regions which are highly divergent among genes from different genotypes and conserved within genes of VP4s in same genotypes were designed. Then four sets of primers for PCR amplified DIG labeled probes were designed and corresponding DIG-labeled specific P genotype probes were synthesized with PCR by using VP8* genes of Beijing field HRV strains representing P-genotypes P[8]a, P[8]b, P[4] and P[6], respectively, as templates. Dot-blot hybridization was developed based on cDNA of VP4 genes. The dot-blot hybridization assay for P genotyping was reliable which was confirmed by sequencing of RT-PCR products of VP4 genes amplified from corresponding clinical samples. P genotyping for VP4 genes from 88 HRV positive specimens from the Outpatient Department (55%, 88/160) and 79 HRV positive specimens from the hospitalized (70.5%, 79/112) children with diarrhea indicated that P[8] a subtype was still the most prevalent sub-genotype, which was 96.6% (85/88) and 62.0% (49/79) respectively. The positive rate for P[8]b subtypes in hospitalized children with HRV diarrhea was higher (27.9%, 22/79) than that of in outpatient (2.3%, 2/88) HRV infected children. HRV with P[4] genotype was only found in one of the hospitalized children (1.3%, 1/79), and HRV with P[6] genotype was not detected from specimens either from outpatient or inpatient. G9P[8]b was the predominant combination among the P[8]b subtype of HRV positive specimens in this study. The results in this study indicated that G9P[8]b HRV circulated in children with diarrhea in Beijing.
Base Sequence ; Capsid Proteins ; genetics ; isolation & purification ; Child ; China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Genotype ; Humans ; Rotavirus ; genetics ; Rotavirus Infections ; epidemiology ; virology

OBJECTIVETo learn the relationship between severity of rotavirus diarrhea and serotype G and genotype P.

METHODThe clinical information and fecal specimens of hospitalized children less than 5 years of age with acute diarrhea in four sentinel hospitals were collected from Aug 2001 to July 2003. Specimens were tested and typed for rotavirus. Each child with rotavirus infection was assessed for severity of diarrhea according to the 20-points scoring system of Vesikari.

RESULTSWhen combined with P[8], the severity scores for rotavirus diarrhea of P[8]G1 and P[8]G3 were 13 and 12 points, respectively, and the durations of diarrhea were 6 days and 5 days, respectively. The percentage of fever in patients with diarrhea caused by P[8]G1 was higher than that in those with diarrheas caused by P[8]G3 (97 percent vs. 73 percent). And the highest temperature in the cases with diarrheas caused by G1 and G3 was 39 degrees C and 38.6 degrees C, respectively. When combined with G3, the difference of diarrhea severity scores between P[4]G3 and P[8]G3 was not significant. But duration of diarrhea caused by P[4] was longer than that of diarrheas caused by P[8] (6.5 days vs. 5 days) and the maximum times of vomiting in P[8] cases was higher than in p[4] cases (4 times vs. 3 times per day). There was no significant difference in other clinical features between P[8] and P[4] infected cases.

CONCLUSIONWhen combined with P[8], RV diarrhea caused by G1 was associated with higher severity scores than diarrhea caused by G3. When combined with G3, there was no significant difference in severity scores between P[4] and P[8] infected cases.

Child, Preschool ; Diarrhea ; complications ; immunology ; pathology ; virology ; Female ; Fever ; etiology ; Genotype ; Humans ; Infant ; Male ; Rotavirus ; genetics ; immunology ; isolation & purification ; Rotavirus Infections ; complications ; immunology ; pathology ; virology ; Severity of Illness Index

Abstract:This study aims to investigate the genetic characteristics of group A rotavirus (GARV) G9P[8] strains from infantile diarrhea samples in Hebei Lulong region from 2009 to 2011. We randomly selected five GARV G9P[8] strains in Hebei Lulong region from 2009 to 2011, amplified the 11 gene fragments of GARVs by RT-PCR, and analyz their full-genome sequences by homology and phylogenetic analysis with DNAStar and MEGA. The nucleotide homology between strains LL11131077 and LL11131083 in 2011 was significantly higher than hat etween them and the other three strains in 2009 and 2010. The G9P[8] GARVs circulating in Hebei Lulong region from 2009 to 2011 elenged to the same genotype as the prevalent G9P[8] GARVs in other parts of the world. However,the two strains in 2011, compared with those in 2009 and 2010, were located in a different sub-branch of the phylogenetic tree and had amino acid mutations at many sites.
China ; Feces ; virology ; Genome, Viral ; Genotype ; Humans ; Molecular Sequence Data ; Phylogeny ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; virology ; Viral Proteins ; genetics

Group A rotavirus is one of the most significant etiological agents which causes acute gastroenteritis among infants and young children worldwide. So far, several method which includes electron microscopy (EM), enzyme immunoassay (EIA), reverse transcription-polymerase chain reaction (RT-PCR)and Real-time Quantitative PCR has been established for the detection of rotavirus. Compared with other methods, Real-time quantitative PCR have advantages in specificity, sensitivity, genotyping and quantitative accuracy. This article shows a overview of the application of real-time quantitative PCR technique to detecte group A rotavirus.
Animals ; Humans ; Real-Time Polymerase Chain Reaction ; methods ; trends ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; diagnosis ; virology ; Viral Proteins ; genetics