Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 2 de 2
Filter
Add filters








Language
Year range
1.
Article in Chinese | WPRIM | ID: wpr-511709

ABSTRACT

Objective To investigate the natural infection of Theiler's murine encephalomyelitis virus (TMEV) in mice,and to survey the distribution of virus in tissues and the changes of serum antibody in the experimentally TMEV-infected mice.Methods Enzyme linked immunosorbent assay (ELISA) and fluorescence quantitative RT-PCR (qRT-PCR) assay were used to detect the antibody and nucleic acid of TMEV in clinical samples.These samples included SPF mice collected from Guangdong area in 2010-2015,mice obtained from a non-barrier laboratory rodent colony,and wild Rattus norvegicus live-trapped around the non-barrier laboratory rodent colony.36 ICR mice were intracerebrally inoculated with TMEV BeAn strain.The clinical signs of the animals were observed daily post-inoculation.Three mice were euthanatized at day 0,3,7,10,17,21,31,39 and 46 post-inoculation (dpi),respectively.Tissue and serum samples were collected for TMEV detection.Results The TMEV antibody and nucleic acid positive rates of SPF mice collected from Guangdong area in 2010-2015 were 5.29% (n=2834) and 27.27% (n=457),respectively.The TMEV antibody and nucleic acid positive rates of the mice obtained from a non-barrier laboratory rodent colony were 71.95% (n=82) and 53.66% (n=82),respectively.The TMEV nucleic acid positive rate of wild Rattus norvegicus was 25.93% (n=27).In the TMEV positive mice,only two mice showed obvious clinical symptoms.The cecal contents,feces and brain samples were the best candidates for qRT-PCR assay.The viral nucleic acid could be detected in the brain,heart,liver,lung and stomach of ICR mice at 3 dpi,but no viral nucleic acid was detected in the spleen,kidney,and cecum.The viruses in liver,heart,lungs and stomach were completely cleared at 10 dpi,and the viruses persisted in the brain throughout the experiment.The TMEV antibody could be detected at 7 dpi,and then the antibody positive rate reached 100% at 17 dpi.The antibody level increased gradually and maintained up to 46 days.ICR mice showed latent infection after TMEV inoculation,with no obvious symptoms and eye pathological changes.Conclusions The experimental mice and wild Rattus norvegicus in Guangdong area are both infected with TMEV,and the infection rate is high.The mice inoculated with TMEV BeAn strain show latent infection.The TMEV antibody produced in mice can be detected at 7 dpi and persisted until the end of the experiment.The viruses are found in the liver,heart,lung and stomach for a short time,but are persisted in the brain for a long time.There is a good consistency of TMEV detection between qRT-PCR and ELISA.The qRT-PCR assay can be used as a powerful complement method for the national standard of laboratory animals.

2.
Article in Chinese | WPRIM | ID: wpr-467218

ABSTRACT

Objective To establish a rapid,specific and sensitive TaqMan real-time fluorescence quantitative PCR assay for detection of murine polyomavirus ( MPyV) .Methods The specific primers and TaqMan probe were designed based on genome sequence of MPyV.The primers amplified a 69 bp fragment.After optimizing the reaction system and reaction condition, the standard curve was plotted by detecting recombinant plasmid standards.The specificity, sensitivity and reproducibility of this method were evaluated.In addition, samples of lungs, spleens and feces obtained from experimentally infected mice and 86 clinical samples were used to validate the efficacy of this real-time PCR assay.Results The specificity assay showed that this assay could specifically detect MPyV and the sensitivity for MPyV was about 100 copies/well.The coefficients of variation ( CV) of both intra-assay and inter-assay were less than 1.13%.All of the samples from experimentally infected mice were positive for MPyV and 3 out of 86 clinical samples were positive by this TaqMan-PCR detection with a positive rate of 3.5%.Conclusions The real-time fluorescence quantitative TaqMan-PCR assay established in this study has high specificity, sensitivity and stability.It can be used for clinical diagnosis, routine detection and epidemiological investigation of murine polyomavirus infections.

SELECTION OF CITATIONS
SEARCH DETAIL