应用多重PCR方法检测石蜡包埋组织标本中的分枝杆菌DNA

应用多重PCR方法检测并鉴别石蜡包埋组织中的结核分枝杆菌复合体与非结核分枝杆菌DNA扩增片段类型 ,为结核分枝杆菌复合体感染与非结核分枝杆菌感染的病理学诊断提供一种补充的鉴别诊断方法。应用三对具有特异性的寡核苷酸引物 ,进行多重PCR扩增。这三对引物分别对应于分枝杆菌 6 5kD表面抗原、结核分枝杆菌插入序列IS6 1 1 0及人类β 珠蛋白基因的部分序列 ,其扩增产物分别为 3 83bp、1 2 3bp和 2 6 8bp。此种多重PCR方法检测的灵敏度为 0 6pg。经多重PCR扩增后进行凝胶电泳 ,结核分枝杆菌复合体 (结核分枝杆菌、牛型结核分枝杆菌、BCG)均可见 3 83bp、1 2 3bp片段 ,而非结核分枝杆菌 (鸟、龟、瘰疬、蟾蜍、堪萨斯、胞内、耻垢分枝杆菌 )仅见 3 83bp片段 (猿猴分枝杆菌与结核分枝杆菌复合体相同 )。与上述相比 ,分枝杆菌感染的临床标本分别增加了一条 2 6 8bp片段。对 2 0 9例临床初步诊断为淋巴结结核病人的石蜡包埋组织标本进行了多重PCR检测 ,1 93例病理诊断为淋巴结结核、结核性肉芽组织、结核性肉芽肿性炎症病人的标本 ,检测结果符合结核分枝杆菌复合体感染。1 6例病理诊断为可疑淋巴结结核病人的标本 ,1 5例检测结果符合结核分枝杆菌复合体感染 ,1例符合非结核分枝杆菌感染。此种多重PCR方法可?

重组分枝杆菌噬菌体检测分枝杆菌的发光研究

采用生物发光方法检测重组的分支杆菌噬菌体对不同细菌的发光反应 ,并比较了仅在特定的温度范围内才能繁殖的温敏噬菌体Phage 88和正常噬菌体Phage 40对分枝杆菌感染活力测定时发光强度的差异 ,以建立用不同类型重组噬菌体检测结核分枝杆菌耐药性的方法和条件 .结果显示两种噬菌体对各种分枝杆菌作用后均有发光 ,对非分枝杆菌发光值很低 ,两者差异有显著性 ;不同的分枝杆菌发光值有差异 :卡介苗的发光值最高 ,结核分枝杆菌的发光值最低 ;温敏噬菌体Phage 88的检测灵敏度大于正常噬菌体Phage 40 ,差异显著 .因此可认为两株噬菌体均可特异地检测结核分枝杆菌 ,但Phage 88的效果优于Phage 40 .

外周血白细胞中结核分枝杆菌DNA检测结果与肺结核类型关系

目的：探讨外周血白细胞中结核分枝杆菌DNA检测结果与肺结核类型的关系，以便为临床正确选择检测标本，从而提高检出率提供依据。方法：应用聚合酶链反应技术，同时检测110例Ⅰ-Ⅳ型肺结核患者的外周血白细胞及痰液标本中结核分枝杆菌。结果：Ⅰ型、Ⅱ型、Ⅲ型、Ⅳ型肺结核患者白细胞中阳性率分别为88.5%、96.4%、25.8%、8.0%,各型间差异显著（P＜0.05）；痰液标本中阳性率分别为53.8%,60.7%,58.1%,58.1%，各型间无显著性差异（P＞0.05）。结论：检测结核分枝杆菌DNA时，Ⅰ型和Ⅱ型肺结核宜选用外周血白细胞标本，而Ⅲ型和Ⅳ型肺结核不宜选用外周血白细胞标本，Ⅰ-Ⅳ型肺结核均可选用痰液标本。

社区获得性快生长型非结核分支杆菌肺病58例临床分析

目的 了解社区获得性快生长型非结核分枝杆菌肺病的临床特点。方法 对 1 986年 1月至1 997年 1 2月间痰抗酸杆菌培养阳性 3 90 7例中 ,经菌型鉴定证明为快生长型非结核分枝杆菌肺病 58例进行回顾性分析。结果 按Runyon分类法 ,感染Ⅳ型偶然分枝杆菌 3 8例 ,龟分枝杆菌 2 0例。男 3 9例 ,女 1 7例 ,病程 8～ 4 0年。主要症状为咳嗽、咳白粘痰、咯血、发热。X线胸片两侧受累者 60 .3 % ,左侧 2 4 1 % ,右侧 1 5.5%。对异烟肼、链霉素、乙胺丁醇、利福平、对氨基水杨酸、丙硫异烟胺耐药率很高 ,对卡那霉素耐药率相对较低。结论 快生长型非结核分枝杆菌肺病病程长 ,症状不特异 ,耐药率高 ,抗结核治疗疗效差。

Pathotype Structure of Magnaporthe grisea in the Fields of Chongyang and Yuan'an in Hubei Province,China

[Objective] This study aimed to explore the pathotype structure of Magnaporthe grisea in Chongyang and Yuan'an in Hubei Province,China.[Method] From the rice-growing fields of Chongyang and Yuan'an in Hubei Province where rice blast occurs frequently,60 isolates which were pathotyped against two sets of host differentials:Chinese host differentials and CO39 NILs,were obtained.Then,20 pathotypes with the six indica host differentials(CO39 NILs) were observed,while 13 pathotypes in four race groups were observed out of the same single spore isolates with Chinese host differentials which consists of three indica cultivars and four japonica cultivars.The diversity of the pathotypes of M.grisea populations tested by CO39 NILs was 2.54 and the pathotype 137.1 occurred at predominantly high frequency(21.67%).The diversity of physiological races of M.grisea populations tested by Chinese host differentials was 1.22 and the race group ZA occurred at predominantly high frequency(73.33%).The diversity of physiological races of M.grisea in Chongyang and Yuan'an were also calculated.Overall,the diversity of pathotypes of M.grisea in Yuan'an was higher than that in Chongyang with the two sets of the host differentials.[Conclusion] This study provided current information on the pathotype spectrum of M.grisea populations in the rice fields of Hubei Province to allow the formulation of viable strategies for blast resistance breeding programs in Hubei Province.

Molecular Cloning of PhoR Sensor Domain from Mycobacterium tuberculosis for Structure-Based Discovery of Novel Anti-Tubercular

The emergence of multidrug-resistant strains （MDR-TB） and extensively drug-resistant strains （XDR-TB） has fuelled the quest for novel drugs and drug targets for its successful treatment. One of the potential candidates as novel TB drug target is the PhoR sensor domain, an extracellular domain of PhoR histidine kinase. PhoR sensor domain is part of the two-component system PhoR-PhoP that senses environmental stimuli and relays the signal to control the expression of 78 virulent associated genes in Mycobacterium tuberculosis. 3D structure of the PhoR sensor domain will facilitate the structure based drug discovery of novel anti- tubercular. In this study, we successfully predicted and isolated the gene encoding PhoR sensor domain from Mycobacterium tuberculosis H37Rv, cloned it in pGEM-T vector and subcloned it in pRSET emGFP expression vector. PhoR sensor domain was successfully cloned and would be used for further expression, purification and crystallization studies.