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1.
Chinese Journal of Virology ; (6): 1-7, 2016.
Article in Chinese | WPRIM | ID: wpr-296225

ABSTRACT

Given the Ebola outbreak in West Africa and the risks of spread to other regions, a rapid, sensitive and simple method for the detection of the Ebola virus (EBOV) is of great significance for the prevention and control of Ebola. We developed a simple colorimetric isothermal multiple self-matching initiated amplification (IMSA) for rapid detection of the Zaire subtype of the Ebola virus (EBOV-Z). This method employed six primers that recognized seven sites of the EBOV-Z nucleoprotein gene for amplification of nucleic acids under isothermal conditions at 63 degrees C for 1 h. Amplification products were detected through visual inspection of color change by pre-addition of hydroxyl naphthol blue dye. Relative sensitivity was validated by detection of serial tenfold dilutions of virus-like particles containing the partial EBOV-Z nucleoprotein gene and mock clinical sample. Specificity of IMSA was validated by detection of the plasma of 30 healthy volunteers, the dengue virus, and Japanese encephalitis virus. IMSA had comparable sensitivity to Reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and cross-reaction with human plasma or other viruses was not observed. Reverse transcription-isothermal multiple self-matching initiated amplification (RT-IMSA) was also evaluated and compared in parallel with the commercial RT-qPCR kit for detection of EBOV-suspected samples of human blood in Sierra Leone. Sensitivity and specificity of the RT-IMSA was 91.4% and 100%, respectively. These data suggest that RT-IMSA is a valuable tool for the detection of the EBOV with the distinct advantages of simplicity and low cost compared with RT-qPCR.


Subject(s)
Humans , Colorimetry , Methods , DNA Primers , Genetics , Ebolavirus , Genetics , Hemorrhagic Fever, Ebola , Diagnosis , Virology , Nucleic Acid Amplification Techniques , Methods
2.
Chinese Journal of Zoonoses ; (12): 216-221, 2015.
Article in Chinese | WPRIM | ID: wpr-460504

ABSTRACT

To identify novel transcripts and sRNA in genome of B .melitensis by transcriptome sequencing ,total RNA were extracted from B .melitensis culture and rRNA were removed .After the addition of adaptor ,RNA was reversely transcribed into cDNA ,which were then subjected to PCR amplification and sequencing .The generated reads were mapped to genome se‐quence of B .melitensis strain 16M .With the mapping results ,novel transcripts and sRNA were identified by bioinformatics methods .Sequencing results analysis showed that genome sequence was covered with the reads with good quality .A total of 773 genes were extended in their 5′and/or 3′ends of their original locations .Sixteen novel transcripts and 241 sRNAs candi‐dates were identified .RT‐PCR showed that some of the sRNAs were differentially expressed under stress conditions .In B . melitensis genome ,there is novel transcript which is not predicted .The sRNA does exist in B .melitensis and were expressed under different conditions .

3.
Journal of Environment and Health ; (12)1989.
Article in Chinese | WPRIM | ID: wpr-544159

ABSTRACT

Objective To investigate whether ?OH radical species present in the acid oxidation-potential water (AOW). Methods Electron paramagnetic resonance spectroscopy (EPR) coupled with the spin trapping technique was used for determination of ?OH radicals. Results A seven-line spectrum characteristic of 5, 5-dimethyl-2-pyrrolidone-N-oxyl (DMPOX) was formed at first, at 60 min the spin adducted DMPO-OH with four-line spectrum characteristic and at 120 min, no spectrum was observed. Conclusion ?OH can be detected in AOW at 60 min and it may come from the interaction of active substance in the AOW.

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