Application of polymerase chain reaction-sequence specific primer method in detecting multiple genotypes based on a large sample of mental diseases / 中国组织工程研究

ABSTRACT

BACKGROUND: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is not applicable for clinical fast detection or the detection analysis of multiple genes on a large sample basis for candidate genes of mental diseases, while polymerase chain reaction-sequence specific primer (PCR-SSP) method can make up for these deficiencies.OBJECTIVE: To establish a PCR-SSP method with multiple gene polymorphism, and popularize its application in the molecular study of mental diseases.DESIGN: Observational trial.SETTING: Department of Molecular Biology and Department of Pathophysiology at Xinxiang Medical College.MATERIALS: The experiment was carried out in the Department of Molecular Biology at Xinxiang Medical College from December 2005 to October 2006. A total of 200 blood samples from patients with mental disease were provided by the Second Affiliated Hospital of Xinxiang Medical College, 0.5 mL each sample. All the patients were informed and agreed with the experiment.METHODS: PCR-SSP was used to analyze the polymorphism of the following genes: variation of tumor necrosis factor (TNF)-α-308A/G and -238G/A, variation of interleukin (IL)-6-174G/C, variation of the 188th base pair (C/T) of CYP2D6 *10B exon, variation of the 472nd base pair (G/A) of COMT gene, and variation of the 941st base pair (G/T) of the CDNA sequence of the 8th exon of MAOA gene.MAIN OUTCOME MEASURES: Gene polymorphism of blood samples analyzed with PCR-SSP.RESULTS: The detected genotypes for TNF-α-308NG included G/G and A/G; the detected genotypes for TNF-α-238G/A included G/A, G/G and A/A; the detected genotypes for IL-6-174G/C were all G/G homozygotes; the detected genotypes for the 188th base pair (C/T) of CYP2D6*10B exon included C/C, C/T and T/T; the detected genotypes for the 472nd base pair (G/A) of COMT included A/G, A/A and G/G; the detected genotypes for the 941st base pair (G/T) of CDNA sequence of the 8th exon of MAOA included T/G, T/T and G/G.CONCLUSION: PCR-SSP is suitable for the polymorphism analysis of the mononucleotide site variation of multiple genes on a large sample basis, which is fast and accurate with low cost and is worthy to be popularized in the molecular study on mental diseases.