基因多态性蕴藏着大量遗传信息,分型技术是研究多态性的主要研究手段。本文建立了一个基于连接酶检测反应的基因多态性并行检测系统,该系统应用连接酶检测反应与基因芯片技术。对AGT M 235T 3种基因型和ACE I/D 3种基因型分型,同直接测...基因多态性蕴藏着大量遗传信息,分型技术是研究多态性的主要研究手段。本文建立了一个基于连接酶检测反应的基因多态性并行检测系统,该系统应用连接酶检测反应与基因芯片技术。对AGT M 235T 3种基因型和ACE I/D 3种基因型分型,同直接测序分型结果一致。将该系统应用于168份临床样本,经过统计学分析,用卡方检验发现高血压组和正常组AGT M 235T多态性存在显著差异(2χ=6.191,P<0.05),但ACE I/D多态性没有显著差异(χ2=5.241,P>0.05)。展开更多
The genes for 5-1ipoxygenase activating protein （ALOX5AP） and phosphodiesterase 4D （PDE4D） have been demonstrated as susceptibility genes for lacunar in the Icelandic and Pakistani populations, but little is known...The genes for 5-1ipoxygenase activating protein （ALOX5AP） and phosphodiesterase 4D （PDE4D） have been demonstrated as susceptibility genes for lacunar in the Icelandic and Pakistani populations, but little is known about the role of these genes in Chinese populations. The present study utilized polymerase chain reaction and ligase detection reaction to detect single nucleotide polymorphisms （SNPs） in 280 consecutive stroke patients and 258 unrelated population-based controls from Nanjing, Jiangsu Province, China. The allele frequency, genotypes, and haplotypes of the two SNPs （rs456009 and rs966221） in PDE4D were similar between the two groups. However, A allele frequency of rs4073259 （A/G） and rs4769055 （A/C） in the ALOX5AP gene exhibited differences in two groups, and especially the haplotype of the SNP was significantly different between the two groups. Results suggested that the ALOX5AP gene might be involved in lacunar infarct, while PDE4D gene was not a risk factor for lacunar infarct in individuals from Jiangsu Province, China.展开更多
AIM: To compare the ligase detection reaction (LDR) and real-time PCR for detection of low abundant YMDD mutants in patients with chronic hepatitis B infection.METHODS: Mixtures of plasmids and serum samples from 52 c...AIM: To compare the ligase detection reaction (LDR) and real-time PCR for detection of low abundant YMDD mutants in patients with chronic hepatitis B infection.METHODS: Mixtures of plasmids and serum samples from 52 chronic hepatitis B patients with low abundant lamivudine-resistant mutations were tested with LDR and real-time PCR. Time required and reagent cost for both assays were evaluated.RESULTS: Real-time PCR detected 100, 50, 10, 1 and 0.1% of YIDD plasmid, whereas LDR detected 100, 50, 10, 1, 0.1, and 0.01% of YIDD plasmid, in mixtures with YMDD plasmid of 106 copies/mL. Among the 52 clinical serum samples, completely concordant results were obtained for all samples by both assays, and 39 YIDD, 9 YVDD, and 4 YIDD/YVDD were detected. Cost and time required for LDR and real-time PCR are 60/80 CNY (8/10.7 US dollars) and 4.5/2.5 h, respectively.CONCLUSION: LDR and real-time PCR are both sensitive and inexpensive methods for monitoring low abundant YMDD mutants during lamivudine therapy in patients with chronic hepatitis B. LDR is more sensitive and less expensive, while real-time PCR is more rapid.展开更多
目的：探讨人类白细胞抗原（ HLA）-DQA1位点基因多态性与尘肺病发病的相关性。方法采用1∶1配比的病例-对照研究方法，选取130例已确诊为尘肺病的患者作为尘肺组，另选130例与尘肺组年龄和累计接尘工龄相近、同性别、同民族、同一工作...目的：探讨人类白细胞抗原（ HLA）-DQA1位点基因多态性与尘肺病发病的相关性。方法采用1∶1配比的病例-对照研究方法，选取130例已确诊为尘肺病的患者作为尘肺组，另选130例与尘肺组年龄和累计接尘工龄相近、同性别、同民族、同一工作地点的健康接尘工人作为对照组。用多重聚合酶链反应-连接酶检测反应分型法，检测2组研究对象HLA-DQA1共6个位点的等位基因，并分析其与尘肺病的相关性。结果尘肺组HLA-DQA1＊0201和HLA-DQA1＊0601基因频率均高于对照组（14.62％ vs 8.85％，12.69％ vs 7.31％），而HLA-DQA1＊0101基因频率低于对照组（34.23％vs 43.08％），差异均有统计学意义（P＜0.05）。 Logistic回归分析显示，HLA-DQA1＊0101、HLA-DQA1＊0201和HLA-DQA1＊0601等位基因频率与尘肺病有关联（P＜0.05），优势比分别为0.688、1.764和1.844，95％可信区间分别为（0.482～0.981）、（1.018～3.055）和（1.019～3.336）。结论 HLA-DQA1＊0201和HLA-DQA1＊0601基因可能是尘肺病的易感基因，HLA-DQA1＊0101基因可能是尘肺病的保护基因。展开更多
AIM: To establish a multiple detection method based on comparative polymerase chain reaction (cPCR) and ligase detection reaction (LDR)/ligase chain reaction (LCR) to quantify the intestinal bacterial components. METH...AIM: To establish a multiple detection method based on comparative polymerase chain reaction (cPCR) and ligase detection reaction (LDR)/ligase chain reaction (LCR) to quantify the intestinal bacterial components. METHODS: Comparative quantification of 16S rDNAs from different intestinal bacterial components was used to quantify multiple intestinal bacteria. The 16S rDNAs of different bacteria were amplified simultaneously by cPCR. The LDR/LCR was examined to actualize the genotyping and quantification. Two beneficial (Bifidobacterium , Lactobacillus ) and three conditionally pathogenic bacteria (Enterococcus , Enterobacterium and Eubacterium ) were used in this detection. With cloned standard bacterial 16S rDNAs, standard curves were prepared to validate the quantitative relations between the ratio of original concentrations of two templates and the ratio ofthe fluorescence signals of their final ligation products. The internal controls were added to monitor the whole detection flow. The quantity ratio between two bacteria was tested. RESULTS: cPCR and LDR revealed obvious linear correlations with standard DNAs, but cPCR and LCR did not. In the sample test, the distributions of the quantity ratio between each two bacterial species were obtained. There were significant differences among these distributions in the total samples. But these distributions of quantity ratio of each two bacteria remained stable among groups divided by age or sex. CONCLUSION: The detection method in this study can be used to conduct multiple intestinal bacteria genotyping and quantification, and to monitor the human intestinal health status as well.展开更多
文摘基因多态性蕴藏着大量遗传信息,分型技术是研究多态性的主要研究手段。本文建立了一个基于连接酶检测反应的基因多态性并行检测系统,该系统应用连接酶检测反应与基因芯片技术。对AGT M 235T 3种基因型和ACE I/D 3种基因型分型,同直接测序分型结果一致。将该系统应用于168份临床样本,经过统计学分析,用卡方检验发现高血压组和正常组AGT M 235T多态性存在显著差异(2χ=6.191,P<0.05),但ACE I/D多态性没有显著差异(χ2=5.241,P>0.05)。
基金the General Program of National Natural Science Foundation of China,No.30870125
文摘The genes for 5-1ipoxygenase activating protein （ALOX5AP） and phosphodiesterase 4D （PDE4D） have been demonstrated as susceptibility genes for lacunar in the Icelandic and Pakistani populations, but little is known about the role of these genes in Chinese populations. The present study utilized polymerase chain reaction and ligase detection reaction to detect single nucleotide polymorphisms （SNPs） in 280 consecutive stroke patients and 258 unrelated population-based controls from Nanjing, Jiangsu Province, China. The allele frequency, genotypes, and haplotypes of the two SNPs （rs456009 and rs966221） in PDE4D were similar between the two groups. However, A allele frequency of rs4073259 （A/G） and rs4769055 （A/C） in the ALOX5AP gene exhibited differences in two groups, and especially the haplotype of the SNP was significantly different between the two groups. Results suggested that the ALOX5AP gene might be involved in lacunar infarct, while PDE4D gene was not a risk factor for lacunar infarct in individuals from Jiangsu Province, China.
文摘AIM: To compare the ligase detection reaction (LDR) and real-time PCR for detection of low abundant YMDD mutants in patients with chronic hepatitis B infection.METHODS: Mixtures of plasmids and serum samples from 52 chronic hepatitis B patients with low abundant lamivudine-resistant mutations were tested with LDR and real-time PCR. Time required and reagent cost for both assays were evaluated.RESULTS: Real-time PCR detected 100, 50, 10, 1 and 0.1% of YIDD plasmid, whereas LDR detected 100, 50, 10, 1, 0.1, and 0.01% of YIDD plasmid, in mixtures with YMDD plasmid of 106 copies/mL. Among the 52 clinical serum samples, completely concordant results were obtained for all samples by both assays, and 39 YIDD, 9 YVDD, and 4 YIDD/YVDD were detected. Cost and time required for LDR and real-time PCR are 60/80 CNY (8/10.7 US dollars) and 4.5/2.5 h, respectively.CONCLUSION: LDR and real-time PCR are both sensitive and inexpensive methods for monitoring low abundant YMDD mutants during lamivudine therapy in patients with chronic hepatitis B. LDR is more sensitive and less expensive, while real-time PCR is more rapid.
文摘目的：探讨人类白细胞抗原（ HLA）-DQA1位点基因多态性与尘肺病发病的相关性。方法采用1∶1配比的病例-对照研究方法，选取130例已确诊为尘肺病的患者作为尘肺组，另选130例与尘肺组年龄和累计接尘工龄相近、同性别、同民族、同一工作地点的健康接尘工人作为对照组。用多重聚合酶链反应-连接酶检测反应分型法，检测2组研究对象HLA-DQA1共6个位点的等位基因，并分析其与尘肺病的相关性。结果尘肺组HLA-DQA1＊0201和HLA-DQA1＊0601基因频率均高于对照组（14.62％ vs 8.85％，12.69％ vs 7.31％），而HLA-DQA1＊0101基因频率低于对照组（34.23％vs 43.08％），差异均有统计学意义（P＜0.05）。 Logistic回归分析显示，HLA-DQA1＊0101、HLA-DQA1＊0201和HLA-DQA1＊0601等位基因频率与尘肺病有关联（P＜0.05），优势比分别为0.688、1.764和1.844，95％可信区间分别为（0.482～0.981）、（1.018～3.055）和（1.019～3.336）。结论 HLA-DQA1＊0201和HLA-DQA1＊0601基因可能是尘肺病的易感基因，HLA-DQA1＊0101基因可能是尘肺病的保护基因。
文摘AIM: To establish a multiple detection method based on comparative polymerase chain reaction (cPCR) and ligase detection reaction (LDR)/ligase chain reaction (LCR) to quantify the intestinal bacterial components. METHODS: Comparative quantification of 16S rDNAs from different intestinal bacterial components was used to quantify multiple intestinal bacteria. The 16S rDNAs of different bacteria were amplified simultaneously by cPCR. The LDR/LCR was examined to actualize the genotyping and quantification. Two beneficial (Bifidobacterium , Lactobacillus ) and three conditionally pathogenic bacteria (Enterococcus , Enterobacterium and Eubacterium ) were used in this detection. With cloned standard bacterial 16S rDNAs, standard curves were prepared to validate the quantitative relations between the ratio of original concentrations of two templates and the ratio ofthe fluorescence signals of their final ligation products. The internal controls were added to monitor the whole detection flow. The quantity ratio between two bacteria was tested. RESULTS: cPCR and LDR revealed obvious linear correlations with standard DNAs, but cPCR and LCR did not. In the sample test, the distributions of the quantity ratio between each two bacterial species were obtained. There were significant differences among these distributions in the total samples. But these distributions of quantity ratio of each two bacteria remained stable among groups divided by age or sex. CONCLUSION: The detection method in this study can be used to conduct multiple intestinal bacteria genotyping and quantification, and to monitor the human intestinal health status as well.
