The study of multiple RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 383-385, 2010.
Article
in Chinese
| WPRIM
| ID: wpr-316891
ABSTRACT
<p><b>OBJECTIVE</b>To establish a multiplex RT-PCR-based reverse dot blot hybridization technique to detect influenza viruses.</p><p><b>METHODS</b>Obtain the HA nucleotide sequences of seasonal influenza H1N1, seasonal influenza H3N2, influenza H1N1 and human avian influenza H5N1 from GenBank. Design primers in conservative district and probes t in high variable region respectively, after analyzing the HA nucleotide sequences of influenza virus through the Vector NTI 9.0. Establish and optimize multiple RT-PCR system by comparing amplification efficiency and specificity at different primer concentrations. Establish the reverse dot hybridization system after optimizing the concentration of probes. To compare the sensitivity and specificity of this technique and the general RT-PCR Method through extracting the viral RNA of the mentioned influenza virus which are to be the reference substance.</p><p><b>RESULTS</b>Successfully establish a multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses. This technique is 100-1000 times more sensitive than gel electrophoresis method, and it has a good specificity.</p><p><b>CONCLUSION</b>Successfully established multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Virology
/
Sensitivity and Specificity
/
Reverse Transcriptase Polymerase Chain Reaction
/
Diagnosis
/
Influenza, Human
/
Influenza A Virus, H1N1 Subtype
/
Influenza A Virus, H3N2 Subtype
/
Influenza A Virus, H5N1 Subtype
/
Genetics
/
Methods
Type of study:
Diagnostic study
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2010
Type:
Article
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