λ噬菌体包装蛋白的制备与λ-DNA的体外包装(简报)

构建基因文库是研究真核基因结构与功能以及人类遗传性疾病分子基础的必经之路。而提取高质量的λ噬菌体包装蛋白以便得到高效率的λ-DNA体外包装,则是构建基因文库成     

地中海贫血的快速产前基因诊断

本研究采用聚合酶链反应(PCR)技术,完成了20例重症β-地中海贫血(β-地贫)及2例重症α-地中海贫血(α-地贫)高危胎儿的产前基因诊断。20倒重症β-地贫高危胎儿中,双重杂合子及纯合子6例,杂合子10例,正常胎儿4例,其中10例已经正常分娩后或流产后基因分析验证,结果与产前诊断完全一致。讨论了三种取材方法及影响PCR的主要因素。PCR技术具有简便快速、早期取材、诊断率高及结果准确可靠等优点,便于推广使用。     

内源性逆病毒癌基因V-erbB突变与白血病发生

目的：证明鸡原始红细胞增多症逆病毒癌基因V—erbB在人类白血病和骨髓增生异常综合征(MDS)急者基因组内的存在。方法：应用V—erbB PCR，V—erbB寡核苷酸(Oligo)原位杂交技术和PCR产物DNA测序，对84例MDS、可疑MDS和49例其他相关血液病进行检到。结果：2例患者骨殖细胞基因组内均存在V—erbB亚基因片段，同源性占99．5％，它们具有相同的限制性酶勿位点，因此可以在Southern印迹杂交条件下发生骨髓细胞C—erbB重排和重排／扩增。白前P1P2PCR产物有420bp，P1P3产物有650bp，而白血病(AL—M4)P1P2PCR产物仅390bp，P1P3PCR因P3无模板序列而无结果。提示从白前发展为白血病，可能发生V—erbB亚基因序列的缺失，即缺失突变。结论：大鼠和人白血病和食管癌等可能均起源于其基因组内存在V—erbB亚基因及其缺失突变。白血病发病中除了上述病因以外，骨髓细胞染色体不稳定性可能起协同作用。     

PCR技术及其在中国人β地中海贫血基因突变类型和产前诊断中的应用

地中海贫血、甲型血友病及杜氏进行性肌营养不良(DMD)是我国常见和危害严重的遗传病,目前尚无满意治疗方法。开展基因诊断及产前诊断是防止患儿出生、取得预防效果、提高人口素质、优生优育的重要措施。本项目在国内最先建立并采用国际上先进的PCR即聚合酶链反应技术,大大减化了以往的基因诊断步骤,使之便于在全国推广。我们应用PCR结合ASO(等位基因特异的寡核苷酸)探针斑点杂交技术,PCR结合限制     

甲型血友病基因诊断的研究

甲型血友病是最常见的遗传性凝血系统疾病,由于凝血因子Ⅷ(FⅧ)基因缺陷所致,为X连锁隐性遗传。发病率为1/4,000～10,000新生男婴,占全部遗传性凝血系统疾病总数的3/4。临床表现为频发的关节及软组织出血,重型患者完全丧失劳动能力,甚至死亡。目前尚无根治办法。因此对该病进行有效的遗传控制,防止患儿出生是十分必要的。血液学和免疫生物学检查可以确定甲型血友病患者,但对携带者的确定和高危胎儿出生前     

Prevalence of CMV in arterial walls and leukocytes in patients with athero sclerosis

Objective To investigate the association of cylomegalovirus with atherosclerosis.Methods The presence of cylomegalovirus (CMV) nucleic acids was demonstrated in carotid and coronary arteries of patients with and without atherosclerosis by polymerase chain reaction (PCR).CMV was detected by PCR in the blood of patients with an d without atherosclerosis.Results 83.3%-86.7% of the samples obtained from atherosclerotic patient arterial wall s were shown to contain viral nucleic acids (CMV), CMV could be found among 6.7 % of patients’ arterial wall without atherosclerosis, significant difference can be found between them (P<0.01).In blood CMV could be found in 42.4% of patients with atherosclerosis, in the control group, only 3% of samples had CMV, P<0.01.Conclusion CMV plays an important role in the pathologic process of the atherosclerosis and the atherosclerotic cerebral infarction.     

Detection of factor Ⅷ intron 1 inversion in severe haemophilia A

Objective Screening the intron 1 inversion of factor Ⅷ (FⅧ) in the population of severe haemophilia A(HA) in China and performing carrier detection and prenatal diagnosis. Methods Using LD-PCR to detect intron 22 inversions and multiple-PCR within two tubes to intron 1 inversions in sereve HA patients. Carrier detection and prenatal diagnosis were performed in affected families. Linkage analysis and DNA sequencing were used to verify these tests. Results One hundred and eighteen patients were seven diagnosed as intron 22 inversions and 7 were intron 1 inversions out of 247 severe HA patients. The prevalence of the intron 1 inversion in Chinese severe haemophilia A patients was 2. 8% (7/247). Six women from family A and 2 from family B were diagnosed as carriers. One fetus from family A was affected fetus. Conclusion Intron 1 inversion could be detected directly by multiple-PCR within two tubes. This method made the strategy more perfective in carrier and prenatal diagnosis of haemophilia A.     

北京汉族群体21号染色体上4个STR位点的遗传多态性

目的研究人类21号染色体上唐氏综合症关键区域内或附近的4个短串联重复序列(D21S1432、D21S2039、D21S1414、D21S1270)在中国汉族人群中的遗传多态性。方法应用Chelex法提取北京地区无血缘关系汉族253份个体血样DNA,聚合酶链式反应(PCR)扩增,非变性聚丙酰胺凝胶电泳或ABI鄄310电泳分型结合测序进行STR分型。结果得到4个STR基因座等位基因频率、各基因座的杂合度、个体识别率、多态信息含量等群体遗传学数据。4个位点基因型分布均符合Hardy鄄Weinberg平衡定律。结论21号染色体上4个STR位点多态性较好,基因型分布符合Hardy鄄Weinberg平衡定律,具有较高的个体识别率,在法医学及唐氏综合征的基因诊断和产前基因诊断研究中具有较高的应用价值。     

SARS患者合并肺炎支原体和肺炎衣原体感染的研究分析

目的：分析比较SARS患中肺炎支原体和衣原体感染的水平。方法：收集87例SARS患、105名血站献血员及116例普通发热患的血清，分别检测其血清中肺炎支原体和衣原体抗体。统计学分析采用卡方(X^2)检验。结果：SARS病例组的肺炎支原体IgG阳性率(80．5％)高于发热组(56．9％)和正常组(47．6％)，lgM在SARS病例组(57．5％)也高于发热组(30．2％)，支原体IgG、IgM同时阳性的比较显示，病例组(51．7％)高于正常组(24．8％)，并有显的统计学意义。衣原体的：IgG和IgM阳性率在病例组(56.3％和24.3％)均高于正常组(37．1％和11．4％)。结论：在SARS患中肺炎支原体和衣原体的感染率处于较高水平，需在SARS治疗过程中注意肺炎支原体或肺炎衣原体的合并感染。并对症治疗。