Study On Rapid Identification Of Actinomycetes Abuot Different Groups

Polymerase chain reaction (PCR) is a rapid technology of creating copies of specific fragments of DNA, which rapidly amplifies a single DNA fragment into millions. The technology of PCR can be used to detect microorganisms quickly as long as the primers are suitable.The PCR technique was used to detect different groups of Actinomycetes rapidly in this Experiment, the results obtained are as follows:1. The 16S rDNA sequences of all the 580 type species of Streptomyces and 50 type strains of other genus from GenBank were analyzed and two characteristic sequences of Streptomyces have been found. Using the sequences, a pair of 16S rDNA genus-specific primers of Streptomyces was designed.2. The 16S rDNA specific primers of Streptomyces, Amycolatopsis, Pseudonocardia, Saccharothrix, Micromonosporaceae and Streptosporangiaceae have been exploring the feasibility of rapid detection methods. An optimization of PCR amplification system and reaction condition was carried out by several main factors as follows: annealing temperature; amount of Taq DNA polymerase; primer concentration and amount of DNA template. The optimization was carried out by single-factor cycle method.3. The results of of using tested strains to determine the specificity of the primers show that all specific bands of each population could be obtained by amplifying 6 kinds of actinomycete 16S rDNA specific primers, moreover there is no corresponding band to be amplified from the other populations of Actinomycetes, which indicates that the 6 kinds of 16S rDNA specific primers all have relatively strong specificity. Therefore, the optimal PCR reaction system and condition could be applied in rapid detection of Actinomycetes, which provides a reference for classification and identification of Actinomycetes.