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Objective To develop a dual real-time fluorescent RT-PCR method for rapid detection of enterovirus(EV)and en terovirus type 71(EV71).Methods Specific primers and probes were designed and the dual real-time fluorescent RT-PCR reaction system was established.The quantitative standard curve was drawn;its sensitivity and precision were evaluated.Feces and throat swab specimens of 109 clinical patients with hand foot and mouth disease were collected and tested by using this method.Then the obtained results were compared with those detected by commercial EV71 PCR kit.Results The relative coefficient(2)of EV and EV71 standard curve established by the dual real-time fluorescent RT-PCR method were both 0.998.Its sensitivity reached 0.5 TCID50/mL for detecting EV and 0.05 TCID50/mL for detecting EV71.The within-run precision for detecting EV and EV71 was ＜3％ and total precision≤4％.The results showed good specificity for the detection of enterovirus and non-enterovirus.In 109 detected clinical samples,84 cases of EV positive samples were detected,in which 56 cases were EV71 positive with the total positive rate of 51.4 ％,which was consistent with the result of simple fluorescent RT-PCR commercialization kit(P=1.000).Conclusion The established dual real-time fluorescent RT-PCR method has high sensitivity and good stability,which has an important significance for early high throughput rapid diagnosis of hand foot and mouth disease.
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Objective To investigate the association between the phosphodiesterase 4D(PDE4D) gene rs153031 polymorphism and the susceptibility of unstable angina pectoris(UAP) in Chinese Han population of Changwu region .Methods The PDE4D gene rs153031 polymorphism was genotyped by Taqman probe in 172 UAP patients(UAP group) ,as well as in gender-and-age-matched 220 subjects without coronary heart disease(CHD)(control group) .Results In this crowd ,there was PDE4D gene rs153031 poly-morphism in patients with UAP and in subjects without CHD .Compared with control group ,frequencies of GG ,GA ,AA genotypes and G allele of rs153031 in UAP group showed no statistical differences (P> 0 .05) .Conclusion In Chinese Han population of Changwu region ,PDE4D gene rs153031 polymorphism shows no association with the susceptibility of UAP .
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Objective To investigate the relationship between serum C-reactive protein(CRP) level ,CRP gene C+1444T poly-morphism and the risk with Acute myocardial infarction (AMI) in Chinese Han population in Sunan region .Methods The CRP gene C+1444T polymorphism was genotyped by Polymerase reaction restriction-fragment length polymorphism (PCR-RFLP) and the serum CRP level was measured by enzyme linked immunosorbent assay (ELISA) between 227 patients with AMI(AMI group) and 161 control subjects .Results No differences were found in genotype distribution between AMI group and controls (CC 82 .38% ,CT 17 .62% ,TT 0% vs 86 .96% ,13 .04% ,0% )(P>0 .05) .The serum CRP level in AMI group was significantly higher than controls(P< 0 .01) .There was no differences in the serum level between any genotypes of the CRP gene C + 1444T (P<0 .05) .Conclusion The CRP gene C+1444T polymorphism is not associate with increased risk of AMI ,and it have no effect with the serum level in Chinese Han population in Sunan region .
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Objective To investigate the possible correlation between the monocyte chemoattractant protein-1 ( MCP-1 ) gene A-2518G single nucleotide polymorphism (SNPs) in the promoter region and acute coronary syndrome (ACS) in Chinese Han ethnic population of Sunan region,Methods This study was conducted with a case-control design in 484 ACS patients including 290 acute myocardial infarction (AMI)patients and 194 patients with unstable angina pectoris (UAP) and 346 control subjects ruled out coronary disease by coronary angiography (control group),including 166 patients with coronary atherosclerosis and 180 subjects without coronary stenosis.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for the detection of the A-2518G polymorphism in MCP-1 gene,and then thefrequency of genetype was statistically analyzed.Results There were AA,AG and GG genotypes of MCP-1 gene A-2518G polymorphism in the ACS group and control group.The two groups could be considered as a genetic equilibrium representative by Hardy-Weinberg equilibrium ( P ＞ 0.05 ).Compared with the control group,the frequencies of AA genotype ( 15.32％ vs.16.12％ ),AG genotype (53.47％ vs.51.86％ ),GG genotype (31.21％ vs.32.02％ ) and G allele genotype (57.95％ vs.57.95％ ) in ACS group were not significantly different ( P was 0.083,0.673,0.821 and 1.00,respectively).Multivariate logistic regression analysis indicated that there was no significant correlation between MCP-1 gene A-2518G polymorphism and ACS regardless of differences in gender,age,smoking,diabetes,TG and LDL-C ( P ＞0.05 ).There was no significant difference in gender and age of ACS onset between two groups ( P ＞ 0.05).There were no significant differences in the frequencies of AA,AG and GG genotypes and G allele genotype among AMI group,UAP group and normal coronary group ( P ＞ 0.05).Conclusions The data shows that MCP-1 gene A-2518G polymorphism is not associated with the risk of ACS in the Chinese Han ethnic population living in Sunan region.