Objective: To understand the genotype distribution and transmission pattern of rubella virus (RuV) circulating in Yunnan Province. Methods: Throat swab samples were collected from rubella outbreaks and sporadic cases in nine prefectures/cities of Yunnan Province from 2011 to 2021. Virus isolation, amplification of target genes and sequence determination were performed on the RuV-positive samples. The genotypes and lineages of Yunnan strains were determined by comparing them with the reference strains, and further phylogenetic analysis was performed with Yunnan strains and strains circulating in other provinces of China during the same period. Results: RuV circulating in Yunnan province during 2011-2021 showed significant genetic diversity, and three lineages, 1E-L1, 2B-L1 and 1E-L2, were detected. Two lineage-switches were also identified, including the conversion of 1E-L1 to 2B-L1 between 2012 and 2013, and the replacement of 2B-L1 to 1E-L2 after 2018. The time of the switches was basically consistent with the outbreak in Yunnan province in 2012 and the time of the rubella reemergence and epidemic between 2018 and 2019. The amino acid sequence of RuV virus strains in Yunnan province was highly conserved, and no important functional regions were changed. Conclusions: The transmission pattern of RuV in Yunnan province is generally consistent with the epidemic trend of RuV in other provinces of China.
Humans ; Rubella virus/genetics* ; Phylogeny ; China/epidemiology* ; Rubella/epidemiology* ; Genotype

Objective: To understand the genotype distribution and transmission pattern of rubella virus (RuV) circulating in Yunnan Province. Methods: Throat swab samples were collected from rubella outbreaks and sporadic cases in nine prefectures/cities of Yunnan Province from 2011 to 2021. Virus isolation, amplification of target genes and sequence determination were performed on the RuV-positive samples. The genotypes and lineages of Yunnan strains were determined by comparing them with the reference strains, and further phylogenetic analysis was performed with Yunnan strains and strains circulating in other provinces of China during the same period. Results: RuV circulating in Yunnan province during 2011-2021 showed significant genetic diversity, and three lineages, 1E-L1, 2B-L1 and 1E-L2, were detected. Two lineage-switches were also identified, including the conversion of 1E-L1 to 2B-L1 between 2012 and 2013, and the replacement of 2B-L1 to 1E-L2 after 2018. The time of the switches was basically consistent with the outbreak in Yunnan province in 2012 and the time of the rubella reemergence and epidemic between 2018 and 2019. The amino acid sequence of RuV virus strains in Yunnan province was highly conserved, and no important functional regions were changed. Conclusions: The transmission pattern of RuV in Yunnan province is generally consistent with the epidemic trend of RuV in other provinces of China.
Humans ; Rubella virus/genetics* ; Phylogeny ; China/epidemiology* ; Rubella/epidemiology* ; Genotype

Rubella virus (RV), a member of the family Togaviridae, can induce apoptosis of host cells in vitro. Protein kinases of the Ras-Raf-MEK-ERK pathway and PI3K-Akt pathway play essential roles in virus multiplication, cell survival and apoptosis. Proteins p53 and TAp63 that bind to specific DNA sequences stimulate Bax in a manner to produce functional pores that facilitate release of mitochondrial cytochrome c and downstream caspase activation. In this review, the molecular mechanisms of RV-induced cell apoptosis, including RV-infected cell lines, pathological changes in cell components and apoptosis signaling pathways are summarized.
Apoptosis ; Humans ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinases ; genetics ; metabolism ; Rubella ; genetics ; metabolism ; physiopathology ; virology ; Rubella virus ; genetics ; physiology

57 rubella virus strains were isolated using Vero cell line or Vero/SLAM cell line from patients' throat swabs during rubella outbreaks and sporadics in 10 provinces of China from 2003 to 2007. Fragments of 1107 nucleotides of E1 genes of the isolates were amplified by RT-PCR, the PCR products were directly sequenced and analyzed. The phylogenetic analysis based on 739 nucleotides showed that out of 57 Chinese rubella virus strains, 55 belong to a distinguish branch of 1E genotype when comparing with 1E genotype rubella strains from other countries, and the other 2 Chinese rubella virus strains belong to 2B genotype. Most of the nucleotide mutations of 57 rubella viruses were silent mutations, and the amino acid sequences were highly conserved. Except one amino acid change (Thr212 --> Ser212) in two rubella viruses at the hemagglutination inhibition and neutralization epitopes, there had no change found at the important antigenic epitope sites of the other rubella viruses. 1E genotype rubella viruses isolated from 10 provinces of China from 2003 to 2007, and two imported 2B genotype rubella viruses from Vietnam suggested that 1E genotype was the predominant genotype in this period of time. The rubella virus genotypes circulated during 2003 to 2007 were different from that circulating during 1979 to 1984 and 1999 to 2002, the rubella prevailed in recent years was mainly caused by 1E genotype rubella viruses with multi-transmission routes.
Genotype ; Mutation ; Phylogeny ; Rubella virus ; classification ; genetics ; isolation & purification ; Time Factors

To analyze the genetic characterization of epidemic rubella virus strains isolated in Liaoning from 2007-2012, a total of 145 rubella virus strains were isolated using Vero/Slam cell line from the patients' throat swabs during rubella outbreaks and sporadics cases in Liaoning Province from 2007 to 2012. Fragments of 945 nucleotides containing 1E gene from 145 rubella virus isolates were amplified by RT-PCR, the PCR products were sequenced and analyzed. Based on the 739 nucleotides of 1E gene, the phylogenetic trees were constructed with 32 WHO rubella reference strains of 13 genotypes downloaded from GenBank and 145 rubella virus strains. The results showed that the 145 rubella virus strains in 2007 -2012 belonged to genotype 1E, nucleotide acids and amino acids similarities were 97.2%-100.0% and 97.6%-100.0%, respectively. Compared to the 1E reference strains(Rvi/ Dezhou.CHN/02, RVi/MYS/01), the nucleotide acids and amino acids similarities were 96.6%-99.2% and 98.2%-100.0%, respectively except for one amino acid change (Val246-Ala246) of RVi/Shenyang. Liaoning. CHN/13.11/13, and Asp262-Asn262 of RVi/Shenyang. Liaoning. CHN/13.11/4 and RVi/Liaoyang. Liaoning. CHN/26. 11/2. there had no change found in the important antigenic epitope sites, the hemagglutination inhibition and neutralization epitopes of the other rubella viruses. All the 145 strains isolated had the same amino acid change (Leu338--Phe338) in E1 protein. These findings suggested that genotype 1E of rubella virus was the predominant genotype in Liaoning province. the rubella prevailed in recent six years was mainly caused by rubella viruses genotype 1E with multi-transmission routes.
Amino Acid Sequence ; China ; epidemiology ; Epidemics ; Genotype ; Humans ; Molecular Sequence Data ; Phylogeny ; Rubella ; epidemiology ; virology ; Rubella virus ; classification ; genetics ; isolation & purification ; Sequence Alignment ; Viral Envelope Proteins ; chemistry ; genetics

Analyze the genetic characteristics of sixteen strains of wild-type rubella viruses derived from Vero cells, Rk13 cells or Vero/slam cells, and isolated from throat samples in Shandong province during 2000-2007. The 1107 nucleotide sequence of nucleoprotein (E1) gene of these isolates were amplified by RT-PCR, and the PCR products were directly sequenced. Comparing with the gene tree that was constructed based on the 739 gene sequences of the WHO reference strains, twelve isolated strains belonged to 1E genotype, one strain belonged to 1F genotype, three strains belonged to 2A genotype. The first strain belonged to 1E genotype was isolated in Shandong province in 2001, then genotype 1E became dominant genotype of wild rubella viruses circulated. The 1E genotype circulated from 2006-2007 was different compared with that circulated from 2001 to 2002, but no significant deviation in temporal and geographic distribution was found. The strain belonged to Genotype 1F was only isolated during 2000 to 2001. The three strains of 2A genotype of rubella viruses were similar to rubella viruses vaccine strain (BRDII). The most nucleotide mutation of rubella viruses among the sixteen strains were nonsense mutation, and the amino acid sequences were highly conservative with no change in important antigen sites. Alike the previous reports, there was the same amino acid mutation in protein E1 at the site of 338 in all of the 1E genotype rubella viruses isolated during 2001- 2007 in Shandong (Leu338 --> Phe338).
Animals ; Cercopithecus aethiops ; China ; epidemiology ; Humans ; Molecular Epidemiology ; Molecular Sequence Data ; Phylogeny ; Rubella ; epidemiology ; virology ; Rubella virus ; classification ; genetics ; isolation & purification ; Vero Cells ; Viral Proteins ; genetics

Measles and rubella virus cause fever rash diseases that are uneasy to differentiate clinically from each other. Specific primers and fluorescence-labeled probes were designed, and a multiplex Real-time RT-PCR with an internal control was developed to simultaneously identify the measles and rubella virus. The multiplex Real-time RT-PCR assay was specific and no false positive or false negative results were found. The sensitivity of the assay was 0.1TCID50/mL and 1TCID50/mL for measles and rubella virus respectively. Analysis with 0.1-10(3)TCID50/mL measles or rubella virus samples demonstrated high validity and reproducibility with the coefficient of variability(CV) of below 0.9% for both measles and rubella virus. Using ribonuclease P (RNase P) as internal false negative control, the developed multiplex Real-time RT-PCR assay is suitable for rapid clinical diagnosis of measles and rubella virus.
Fluorescent Dyes ; Humans ; Measles ; diagnosis ; virology ; Measles virus ; genetics ; RNA, Viral ; genetics ; isolation & purification ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Ribonuclease P ; genetics ; Rubella ; diagnosis ; virology ; Rubella virus ; genetics ; Sensitivity and Specificity

OBJECTIVETo understand the hereditary and variant characteristics of rubella virus(RV), especially the strains isolated from China, investigating the epidemical trend and variation principle of RV.

METHODSThe envelope glycoprotein E1 gene was amplified from rubella virus by RT-PCR. After sequencing, the gene sequence was handled by the software DNASTAR and the phylogenetic tree was drawn to analyze the molecular epidemiological characteristics of RV.

RESULTSThe sequence of RV strain JR23 was sequenced, the phylogenetic tree was drawn taking 30 strains isolated at different times and locations in GenBank, including three strains from China as reference. The regions that encode the peptides which react with the HI antibody and the neutralization antibody were compared to show if there was any amino acid mutation in the sequence. (1) In general, the nucleotide and amino acid sequences of RV were highly conserved. The four strains isolated from China had relatively large variations. Strain 379 and strain BRD constituted genotype II, which is different from the other 29 strains. Further study is needed to understand their heritable resources and biological characteristics. (2) Strain JR23 showed little difference from the strains that were epidemic during 1960s in UK, USA and Japan, so maybe it is the derivative strain of that in epidemic 1960s. But the accurate epidemic time is not known.

CONCLUSIONThere are differences among areas and time in epidemics of rubella. The mobility and the region difference seem to be the key factors that affect the epidemic characteristics of RV.

Amino Acid Sequence ; Amino Acid Substitution ; Base Sequence ; China ; epidemiology ; Genotype ; Molecular Epidemiology ; Mutation ; Phylogeny ; Rubella ; epidemiology ; virology ; Rubella virus ; classification ; genetics ; Sequence Analysis, Protein ; Sequence Homology ; Viral Envelope Proteins ; genetics

Throat swabs collected from patients whose serum was measles IgM negative and rubella IgM positive during 2001-2011 were used to conduct cell culture for rubella virus. After identification of cell culture with RT-PCR, nucleotide of gene E1 of rubella virus was amplified and sequenced, followed by molecular epidemiological analysis. A total of 31 rubella viruses were isolated from 60 throat swabs. Compared 27 isolates with the WHO reference strains of all genotypes, phylogenetic tree was constructed based on the amplified 739 nucleotide fragment. These isolates belonged to two different genotypes respectively. Isolates 11009, 11052 and 11106 in 2011 belonged to genotype 2B, and others belonged to genotype 1E. Most of mutations were nonsense mutation, and sequence of amino acid was highly conserved. Amino acid sequence of most isolates of genotype 1E was identical, which suggested rubella viruses from same transmission chain might be transmitted continually since 2001. Rubella virus genotype 2B was found to be popular for the first time in Shanghai in 2011. The nucleotide sequences of these genotype 2B isolates showed 99% identity compared with that of isolates recently from Vietnam, Japan and Argentina. The resources of these strains were not confirmed due to the absence of rubella virus surveillance before.
Adolescent ; Adult ; Amino Acid Sequence ; Child ; Child, Preschool ; China ; epidemiology ; Humans ; Infant ; Male ; Molecular Epidemiology ; Molecular Sequence Data ; Rubella ; epidemiology ; virology ; Rubella virus ; classification ; genetics ; isolation & purification ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; Young Adult

OBJECTIVETo construct a recombinant plasmid vector of the RV specific fragment for expressing the specific fragment of RV E1 protein.

METHODSRNA of the RV attenuated live vaccine Wistar RA27/3 strain was extracted and reversely transcribed. The specific fragment of the E1 gene was amplified and the PCR products cloned in the vector pGEX-2T after purification. Positive clones were selected and identified by two-enzyme digestion and sequence analysis.

RESULTSA 330 bp target fragment was successfully cloned, and the sequence of the recombinant plasmid was consistent with the original sequence.

CONCLUSIONSuccessful cloning of the RV El specific fragment and the construction of the recombinant plasmid have laid a foundation for further expressing the recombinant protein.

Base Sequence ; Cloning, Molecular ; Gene Expression ; Genetic Vectors ; Molecular Sequence Data ; Plasmids ; RNA, Viral ; Reverse Transcriptase Polymerase Chain Reaction ; Rubella virus ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology