Application Of Gene Chip Integrated System To Rapidly Diagnose Eight Pathogens Of Swine Diseases

In this experiment, by taking 8 important pathogens which are affecting the growth, healthy and reproduction in pigs as the research object, a technology based on cDNA microarray assay was established to diagnose these pathogens rapidly at a time. Japanese encephalitis Virus, Porcine pseudorabies virus, Foot -and-Mouth Disease type O and type AISAâ… , Porcine reproductive and respiratory syndrome virus,Porcine parvovirus,Porcine circoviruses and Classical swine fever virus are the research objects.Gene chip technology is a new technology with the development of carrying out human genome project in 90's age of 20 century. It is a rapid, high efficient, paralleled and high throughput diagnostic technology adopting the parallelly processing and highly integrated technology that can detect hundreds of diseases. Biochip technology gradually developed into a laboratory of routine molecular techniques with wide application in scientific research. Compared with the traditional detection methods, it has many advantages, such as mass, high throughput, rapid detection, etc.The aim of this study is to establish a microarray technique to differentiate eight kinds of pathogens at a time in gene level by dealing with the sample at a time. The research contents mainly embrace: eight kinds of specific primers were designed according to conservative region; the extracted nuclear acids were amplified by PCR amplification and the amplified products were inserted into PMDT-18 vector to construct standard templates as probes; the probes were fixed on the amino-modified slides; asymmetric PCR were conducted using nuclear acids of virus to product the target material, and the amplified products were labeled with CY3 and then hybridized with the cDNA probes on the slides. The hybridizing results were scanned by Genepix4000A scanner. The results showed that the amplified product of these eight kinds of viruses could specifically bind to the cDNA on the slide chips, which indicated that these probes could be used to detected viruses specifically; the sensitivity of the test was higher than the common PCR; the results from interassay test were consistent with that from intraassay test, which suggested that the technique to detect these eight kinds of viruses has preferable stability. The results above showed that cDNA chips technique to detect these viruses has a characteristic of high specificity, sensitivity and stability, which will be a novel, reliable method to detect these viruses. As well as disease prevention and control and molecular epidemiological investigations laid the foundation. Therefore, this study has important practical significance and application value.