Identification Of Patients And Carriers Of Spinal Muscular Atrophy By Fluorescent Quantitative Real Time PCR Technology Information Network Management Solution Based On LDAP

I. Identification of patients and carriers of spinal muscular atrophy by fluorescent quantitative real time PCR technologyBackgrounds: Spinal Muscular Atrophy (SMA) represents the second most common autosomal recessive disease in live births. The clinical characteristics of the disease include degeneration of anterior horn cells of the spinal cord, muscular palsy, atrophy, patients often die in respiratory muscle palsy with severe phenotype. The deletion of SMN1 will result in the SMA. SMN2 is highly homologous with SMN1, the mainly difference is a samesense mutation of C→T in exon 7, which indicated about 70% SMN2 mRNA composed dysfunctional protein. As there are no effective therapy method and the highly heterozygosity frequency of 1 in 40, rapidly and accurately detect the carriers and patients is the key to decrease the prevalence of this disease. The common gene diagnosis methods can only detect patients of SMA but not carriers.Objective: To set up a patients and carriers detection method based on the fluorescent quantitative real time PCR technology. Using this method detect 17 SMA families and 15 normal controls.Methods: We developed a novel, rapidly and stable gene diagnosis method based on the quantitative real time PCR technology with specific Taqman and MGB probes, using ALB gene as internal reference, optimized the reaction condition and instituted a standard detect protocol. Through setting up the reaction standard curves, calculating the ratio of standard DNA with sample DNA, than got the copy number of SMN1 in sample DNA.Results: Until now, we have detected 17 SMA families and 15 unrelated normal controls with 75 samples in total, the range of single copy and double copies are within the ranges[0.38, 0.62],[0.78,1.2], respectively. In all of 17 SMA families, there are 40 SMA carriers and 17 SMA patients, no exon 7 deletion were detected in 3 samples, no deletions were detected in all of 15 normal controls. By using PCR-enzyme method, 17 SMA patients were detected, and other 43 samples and 15 normal controls were not discriminated.Conclusion:①Based on the quantitative real time PCR technology, Taqman and MGB probes, ALB gene as internal reference, a novel SMA patients and carriers gene detection method has established. This method is rapidly and accurately.②40 carriers, 17 patients and 3 no deletions were detected by real time method in 17 SMA families with 60 samples in total. No deletions were detected in 15 normal controls.③All of 60 samples of SMA families were detected by PCR-restrictive method and discriminated 17 patients. No deletions were detected in 15 normal controls, which was accordant with results above. II. Information network management solution based on LDAPBackgrounds: Recently, most of the medical research department and general hospital have implemented computer information management, but a lot of problem is still waiting to be resolved. Problems included: storage sub system's pressure increasing fast as the critical process needs more powerful engines; great challenge in system management and data secure because of more and more computer virus and hacker invading; the over set user authority result in lower security, over risk and more management cost.Objective: To design a novel information network management system, to solve the limitation of current management system.Methods: Based on light-weight directory access protocol (LDAP), and advanced computer technologies including Windows 2003, Windows Server Update Service, HP Array management Utility, Firewall, Antivirus software to solve existed problems.Results and Conclusion: The new system has new architecture, and we got better performance in security, stability, remotely and compatibly. Some new characteristics were added. The system has run in the National Laboratory of Medical Genetics for 33 months.