Aug. 12, 2009
The National Microbiology Laboratory (NML) has developed a highly sensitive toolkit to identify orthopoxvirus and distinguish human smallpox from monkey smallpox camel smallpox and vaccines. It contains primers, amplification solution for DNA strand polymerization, positive control, confirming that the test is conducted under conditions conducive to polymerization, and establishing synthetic models to produce genetic material corresponding to HA and cmB gene sequences of human smallpox and monkey smallpox viruses.
The length polymorphism of amplified DNA restriction fragment (PCR-RFLP) was analyzed by NML kit to identify orthopoxvirus. The strands of DNA molecules are isolated and used as a matrix to produce the same DNA molecules as the starting molecules. The NML kit contains primers capable of matching and elongating specific regions (HA or crmb regions) in the DNA strand of the orthopoxvirus. In the presence of orthopoxvirus, appropriate primers bind and produce DNA strands. The obtained genetic material is then subjected to restriction endonucleases (Sau3AI and SPE I of the HA region, and DrI, Alw44I, SSP I and HPA I for crmb), which interact with specific fragments of the genetic material and fragment the chain.
Through the analysis of the enzyme sections, we can identify the existence of orthopoxvirus strains.
Smallpox detection technology is an important tool in the bioterrorism toolkit. The National Microbiology Laboratory kit detects smallpox virus and distinguishes it from other orthopoxviruses, and includes a positive control to verify that test conditions produce reliable results. This is a terrible weapon in smallpox control. I don't think RFLP technology is suitable for field use.
We need smallpox control tools to address the threat of bioterrorism. It is important to quickly identify infections so that rapid action can be taken to avert epidemics. However, in medicine, formal identification of the virus remains difficult, as smallpox is easily confused with chickenpox and is almost impossible to distinguish from smallpox in monkeys, whose pathogens also belong to the genus Orthopoxia. Many orthopox testing techniques are incapable of distinguishing between species or lack of positive controls, and it is not possible to determine whether the test was conducted under the correct conditions. A method of identifying positive smallpox, distinguishing human smallpox from monkey smallpox, camel smallpox and vaccine, supplemented by a positive control, will be an important tool for preventing smallpox epidemics.
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establishing synthetic models
cmb gene sequences
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orthopox testing techniques
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