| As the developing of microbiology and molecule chemistry, the appraisal on the pathogeny was not localized on the common inspection just like distinguish the form and the frame or its physiological characteristic. The molecule biology as a tool was used in the research on the bacterial structure of nucleic acid and its other parts. In recent years bacterial ribosome RNA was found to be the sign of its living. It was implied the current infection if the bacterial rRNA was shown by detected, so the rRNA gene was under intensive study. The structure of the 16S rRNA nucleotide was highly conservative and the length of the fragment comprise inheritance information is adequate. There were variations in different families,genus and species, but the conservations were relative. These characters might be used in the appraisal on bacterium and offer a new gene-diagnose method to the infection disease induced by bacteria. The traditional bacterial culture had its weakness of spent longer time and lower positive-rate and didn't suit the need of modern diagnosis and give treatment. The technology of PCR was used as an examing method in clinical especially in the rapid and sensitive diagnosis on pathogenesis. The technology of gene-chip was a new method for gene-diagnosis provided with rapid, high-effect, sensitivity and economy. So this technology was becoming a hotspot in the present time research. To characterize the role of 16S rRNA gene in the use of quick detection and diagnosis gene-chip to bacteria based on its primers of bacterium and special probes for some bacterium, we did following studies.PART ONE AMPLIFICATION AND APPLICATION OF THE 16S rRNA GENEPURPOSE The bacterial genome DNA was expressed and amplified by the primers designed with the 16S rRNA gene of bacteria. The primers were evaluated in its specificity and its sensitivity. The bacterium were examined with the primers. METHODS PCR primers were constructed based on the high conservative regionof the 16S rRNA gene in bacteria. Conditions of the PCR reaction were created and optimized. The DNAs of various bacterium, two kinds of virus and the human leukocytic had been amplified in the same condition to get the products. The methods such as agarose gel electrophoresis were used to evaluate the specificity and the sensitivity of the primers. RESULTS The fragment about 371 bp was seen in the product of the bacteria. But the same fragment wasn't shown in the product of each two kinds virus and the human leukocytic DNA. The primers were checked about sensitivity and the primers could find 10 E.coli. CONCLUSION The PCR primers of the 16S rRNA gene were specific and sensitive to the bacteria amplification. These primers can be used in amplification to bacteria and in examination of gene chip.PART TWO CONSTRUCTION OF THE QUICK DIGNOSIS GENE CHIPPURPOSE Construction of the quick dignosis gene chip based on 16S rRNA gene of bacteria and the gene chip was used to examine the bacterium. METHODS The specific probes of the Gram's positive bacteria, Gram's negative bacteria, Escherichia coli, streptococcus pneumoniae, staphylococcus aureus were composed based on the 16S rRNA gene and the gene chip made up of them were constructed. The hybridization conditions were already optimized according to selecting the best. The specificity and sensitivity of gene chip were tested by inspecting the PCR product of bacteria. RESULTS The quick diagnosis gene chip was constructed successfully. It wasn't only showing one kind of bacteria but distinguishing the different kinds of bacterium after optimized the reaction conditions. CONCLUSION The gene chip was made for examining bacteria and would be applied for clinical diagnosis.PART THREE THE QUICK DIGNOSIS GENE CHIP USING IN CLINICPURPOSE The quick dignosis gene chip was imposed in examining the clinical samples and the foreground of using in clinic was discussed. METHODS The clinical samplies were cultured and appraised with traditional method and applicated by PCR using the primers. The PCR produces were...
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