Use molecular biology technology test of avian H7N9

　　

　　Science: H7N9 is a subtype of avian influenza virus. Influenza virus particles membrane covered by the two type surface glycoprotein, a type of blood cell agglutinin (H), a type of neuraminidase (N), H was divided into 15 subtypes, N is divided into 9 subtypes. All human influenza viruses can cause avian influenza, but not all of the avian influenza virus can cause a human influenza, avian influenza virus, H3, H5, H7, H9 can infect people, wher H5 is the highly pathogenic. H3 occurs in dogs and humans, according to characteristics of influenza virus can be divided into 135 kinds of HxNx subtypes, H7N9 subtype avian influenza virus is one of them. The biological characteristics of the virus, pathogenicity, the propagation force, is not on the basis of analysis and judgment.

　　

　　Now the lab tests are mostly use RT-PCR (reverse transcription polymerase chain reaction), said that the first PCR (polymerase chain reaction), we all know that DNA is composed of ATCG four base pair double stranded form, the PCR method is to find the peculiar to this virus DNA fragment after addition of the enzyme (is an efficient catalyst) and base (the equivalent of raw materials to production of DNA fragments) can detect the degree. As for the RT (reverse transcription) is to put RNA into DNA.

　　

　　Now look at the eastern treasure laboratory testing process:

　　

　　1 biological safety cabinet (or super clean workbench), extraction of RNA full on the inside operation

　　

　　2 masks, prevent gas operated exhaled on extracting RNA pollution

　　

　　3 refrigeration centrifuge, when using, can guarantee the high speed centrifugal RNA solution does not overheat

　　

　　4.YS-RNA Extraction Kit (centrifugal column), the extraction of H7N9 avian influenza virus RNA

　　

　　5 the submission of samples with organic solvent to make the cells burst release of RNA, after the centrifugal filtration, RNA will remain in the membrane on extraction column

　　

　　6 complete extraction solution, the bottom of the tube that little is

　　

　　7 the use of avian influenza H7N9 RT-PCR gel detection reagent box of H7N9 avian influenza virus, kit usually use a special ultra low temperature freezer at -20 DEG C preservation

　　

　　8.PCR amplification, this instrument can expand just to extract RNA millions of times

　　

　　9 staining solution and electrophoresis apparatus. The PCR amplification products are polar, added to the electrophoresis apparatus inside power will move in the agar gel. Some staining solution is carcinogenic, do a good job of protection suggested doing the experiment

　　

　　10 gel imaging system, because the agar gel added staining solution can make RNA staining and detected ultraviolet lamp this machine, and then entered into the computer to get results

　　

　　11 results from left to right are Maker, positive control, negative control, unknown samples. In the 500bp bands were H7 positive samples, in 300bp bands were N9 positive samples, otherwise as negative samples