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China: T?V Rheinland Group RT-PCR Detection Method Approved

Commonground

Senior Moderator
H5N1 bird flu virus detected
2008-6-30

The highly pathogenic H5N1 avian influenza virus this year is again attacked China and other parts of East Asia. In China, Thailand and Vietnam, the virus has proved fatal to humans.

Avian influenza virus (AIV) diameter of about 100 nanometer. The avian flu virus from eight length between 900 to 2,400 single-nucleotide ribosomal RNA negative chain (RNA) components.

All kinds of birds is the spread of the avian flu virus carriers. 2-5% of wild birds carrying the avian flu virus. Most of the avian flu virus is not fatal to birds and will not transfer to humans.

According to the avian influenza virus subtype of the key surface protein "Hemagglutinin" (HA or H) and "glycoside neuraminidase enzyme" (N) of the English name of the first letter naming the virus. Both of which are concentrated in the virus membrane. One of H5N1 avian influenza virus, has been in Asia and Turkey, causing several deaths, but does not seem to find its person-to-person case.

Circulation outbreak of the H5N1 avian influenza virus enough that the development of a highly sensitive, accurate and rapid diagnostic test of necessity. Its importance is reflected not only in infection control and early anti-viral treatment, but also in the food supply chain quickly and effectively control the product. However, cell culture and subsequent hemagglutinin (HA or H) and neuraminidase enzyme glycoside (N) subtype serology tests require 1-2 weeks to get the test results, therefore, their relatively short shelf life of food Products in the health status assessment of little use. On the other hand, commercial general of the avian flu virus antigen rapid test Although both quick and easy, but subtypes (H1 and the H5 strain of) virus exists cross-reaction, difficult to subtype confirmed. Real-time RT-PCR technique in comparison to other detection methods of detection in the performance of the major advantages of speed and can carry out the analysis of viral load, has a higher sensitivity and specificity. Therefore, the application of real-time RT-PCR technology in molecular biology detection method is the most effective and most rapid of the avian flu virus detection programme.

German T?V Rheinland Group of Shenzhen China's food laboratory directly against the H5N1 avian influenza virus has developed a fast, high sensitivity and specificity, to fluorescent light detection-based, quantitative RT-PCR detection method. Testing methods developed in line with the World Health Organization's avian influenza virus testing standards, test results have the authority.

http://translate.google.com/transla...html&hl=en&ie=UTF8&sl=zh-CN&sl=es&tl=en&tl=en
 
Re: China: T?V Rheinland Group RT-PCR Detection Method Approved

See also in SEL. RES. ABSTRACTS thread #1:

(1.10): J Virol Methods. 2008 Jun 20. [Epub ahead of print]

Development of reverse transcription loop-mediated isothermal amplification for rapid detection of H9 avian influenza virus.


Chen HT, Zhang J, Sun DH, Ma LN, Liu XT, Cai XP, Liu YS. - Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.

Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a unique gene amplification method that can be completed within 45min at 63 degrees C.
In this study, RT-LAMP was used to develop a rapid and sensitive laboratory diagnostic system for the H9 subtype of avian influenza virus (AIV).
The experiment results from the reference strains demonstrated that the established RT-LAMP sensitivity was 10-fold higher than that of RT-PCR, with the detection limit of 10 copies per reaction, and no cross-reactivity was observed from the samples of other related viruses including H5N1, H3N2 subtype of AIV and Newcastle disease virus.
Furthermore, a total of 112 clinical samples were tested by RT-LAMP, RT-PCR, and virus isolation, respectively.
All of the 85 positive specimens identified by virus isolation were also positive by RT-LAMP, while 7 of these samples were missed by RT-PCR.
These results suggest that the present RT-LAMP system may provide a new avenue for the recognition of H9 subtype virus, and may be employed to screen for potential carriers in wild and domestic birds.
PMID: 18572258 [PubMed - as supplied by publisher]
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