Detected Pseudorabies Virus And The Three Major Pathogenic Bacteria Of Microarray Technology

With the development of molecular biology techniques, many molecular biology methods in the diagnosis of animal diseases has been applied. Microarray technology as a high-tech,in the area of small carriers such as slides can be realized simultaneously detect multiple target genes. It is based on PCR, and nucleic acid hybridization principles, prior to the carrier surface of a specific treatment, the oligonucleotide or cDNA fragment in an orderly, high-density land arranged in solid phase carrier to form a DNA microarray, with high precision degree of laser confocal fluorescence detection system for detection of hybridization signals. The advantage of high throughput, parallelism, miniaturization, automation, fast and accurate, multi-function and prevent pollution in order to effectively rule out outside interference. Many scholars at home and abroad are engaged in research on gene chip, gene chip technology has undoubtedly animal disease diagnosis and microbiological test provides a rapid and effective detection. This study tested the preparation for detection of porcine pseudorabies virus and several pathogenic gene chips.Pseudorabies virus (PRV) is a disorder caused by porcine reproductive one of the major pathogenic bacteria,the disease to the swine industry has caused tremendous economic losses, the conventional methods of laboratory diagnosis of the etiology most time-consuming, low sensitivity and require high the operating techniques. In this study, the analysis of porcine pseudorabies virus genome structure of the basis of the PRV and the target gene microarray probes for detection of PRV, chip hybridization temperature and time; The gE gene as a target gene, and primers were designed according to the gene probe, amplified by asymmetric PCR gene, the probes immobilized on aldehyde-treated after chip up detection of pseudorabies virus, was constructed PRV pathogen microarray and reached the detection of porcine pseudorabies virus purposes.Clinical diagnosis of pathogenic microorganisms commonly used methods of cultivation techniques, immunoassay, PCR technology, these techniques in clinical diagnosis played a huge role, but let some shortcomings. Cumbersome and time-consuming culture techniques; immune serum specific technical need, rapid detection and identification of intestinal bacteria to timely and effective prevention, treatment and control of intestinal infectious diseases. In order to explore an accurate, rapid detection method, this study attempts to gene chip technology to the field of pathogen detection, use of prokaryotic 16S rRNA gene conservation and variability of the characteristics of the design selection of the one pair of universal primers, in its conservative area design universal primers, probes in the design of its variable region, multi pathogen detection, the true high throughput detection, under the same conditions as Salmonella, Staphylococcus aureus and Escherichia coli, to detection and identification of bacteria The purpose of preparation of the gene chip can detect Salmonella, Staphylococcus aureus and Escherichia coli.