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TaqMan-rRT-PCR检测猪霍乱沙门菌方法的建立和实验室评价 被引量:2
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作者 张景山 李旭 +2 位作者 闫梅英 阚飙 樊粉霞 《中国人兽共患病学报》 CAS CSCD 北大核心 2021年第5期410-415,共6页
目的基于猪霍乱沙门菌血清型特异性基因,建立TaqMan逆转录实时聚合酶链反应(TaqMan-rRT-PCR)检测猪霍乱沙门菌的方法。方法利用基因组序列比对筛选到猪霍乱沙门菌特有的基因SC0358,通过普通PCR及利用多种血清型沙门菌及非沙门菌菌株共计... 目的基于猪霍乱沙门菌血清型特异性基因,建立TaqMan逆转录实时聚合酶链反应(TaqMan-rRT-PCR)检测猪霍乱沙门菌的方法。方法利用基因组序列比对筛选到猪霍乱沙门菌特有的基因SC0358,通过普通PCR及利用多种血清型沙门菌及非沙门菌菌株共计145株,评价该方法的菌株特异性,针对该基因设计TaqMan-rRT-PCR检测方法的引物,优化反应条件,建立针对该靶基因的TaqMan-rRT-PCR检测方法,以纯菌及血液模拟标本RNA为模板进行敏感性检测。结果利用该方法检测26株猪霍乱沙门菌均为阳性,其余菌株扩增均阴性,对纯菌RNA检测中,TaqMan-rRT-PCR的最低检测限度为5 fg/反应,约为10个拷贝/反应。对全血液模拟样品分析中,敏感性达25 cfu/mL。结论以猪霍乱沙门菌保守、特异基因SC0358建立的TaqMan-rRT-PCR方法,能够简便快捷区分猪霍乱沙门菌与其他血清型沙门菌,尤其能够区分与其有抗原式相同的丙型副伤寒沙门菌,此方法为猪霍乱沙门菌感染的快速诊断提供了简便的手段,可用于对猪霍乱沙门菌的早期诊断。 展开更多
关键词 猪霍乱沙门菌 TAQMAN 检测
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油田生产污水低品位热能的回收方案探讨 被引量:1
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作者 张景山 邓雄 +2 位作者 黄建敏 藏薇 隋志成 《节能与环保》 2019年第2期78-79,共2页
随着我国各主要油田的开发已进入高含水期,大部分油田地面采出液的含水率高达90%以上。特别是油井采用热采和电泵采油技术,井口温度达60~80℃,原油处理中大量采用加热分离处理技术,导致地面分离污水的温度至少在40~50℃。这些生产污... 随着我国各主要油田的开发已进入高含水期,大部分油田地面采出液的含水率高达90%以上。特别是油井采用热采和电泵采油技术,井口温度达60~80℃,原油处理中大量采用加热分离处理技术,导致地面分离污水的温度至少在40~50℃。这些生产污水的低品位热能基本上未能得到有效利用或利用率远远不够。这既造成能源浪费,又造成环境热污染。所以回收利用这些低品位热能具有重要的现实意义。本文分析油田地面生产系统的生产污水低品位热源情况,开展生产污水余热资源的回收方案初步设计,以期能够在国内各大油田得到广泛应用,对我国油田企业的低品位能源利用和节能减排起到一定的推动作用。 展开更多
关键词 油田 低品位热能 回收 方案设计 探讨
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Estimation of Local Energy Density in Relativistic Heavy-Ion Collisions
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作者 张景山 马忠彪 高崇寿 《Chinese Physics Letters》 SCIE CAS CSCD 2001年第8期1015-1017,共3页
The energy density for the central region in relativistic heavy-ion collisions can be estimated via the pseudorapidity distribution of transverse energy.The way to estimate the local energy density for the central reg... The energy density for the central region in relativistic heavy-ion collisions can be estimated via the pseudorapidity distribution of transverse energy.The way to estimate the local energy density for the central region in relativistic heavy-ion collisions is proposed,in which only final state particles emitted from the same source are included.The energy density arrived in the NA49 experiments is about 0.9 GeV/fm^(3). 展开更多
关键词 energy. RELATIVISTIC COLLISIONS
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2010年-2017年南京市农村饮用水微生物监测结果分析 被引量:13
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作者 葛明 张景山 +1 位作者 熊丽林 李峻 《中国卫生检验杂志》 CAS 2019年第4期497-499,共3页
目的掌握本市目前农村饮用水微生物污染状况,了解影响农村饮用水微生物污染的因素。方法 2010年-2017年南京市农村地区采集样本1 936份,按GB/T 5750.12—2006 《生活饮用水标准检验方法微生物指标》进行菌落总数和总大肠菌群卫生检测,按... 目的掌握本市目前农村饮用水微生物污染状况,了解影响农村饮用水微生物污染的因素。方法 2010年-2017年南京市农村地区采集样本1 936份,按GB/T 5750.12—2006 《生活饮用水标准检验方法微生物指标》进行菌落总数和总大肠菌群卫生检测,按GB 5749—2006 《生活饮用水卫生标准》进行评价,对南京市农村饮用水存在的问题提供科学依据。结果共收集2010年-2017年农村饮用水监测样品1 936份,合格1 572份,总合格率为81.2%,各年合格率呈上升趋势。出厂水和末梢水2类样品合格率分别为71.4%和85.3%;丰水期和枯水期2类样品合格率分别为77.1%和85.1%;不消毒、复合二氧化氯、高纯二氧化氯、漂白粉和液氯5类样品合格率依次为56.9%、67.6%、70.5%、63.6%和98.2%。结论需进一步扩大区域供水范围,加强农村水厂建设和对其净化、消毒的卫生监督与指导,提高农村饮用水卫生安全,保护公众健康。 展开更多
关键词 饮用水 微生物 农村
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高通量半自动化细菌全基因组序列测定体系的构建
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作者 张景山 龙政 +3 位作者 王紫鉴 肖文静 王群 赵宏群 《中国媒介生物学及控制杂志》 CAS 2020年第6期735-737,共3页
目的建立高通量、半自动化细菌全基因组序列测定体系,满足基于基因组序列的细菌性病原体监测的需求。方法使用自动化液体处理器、可丢弃性机械手枪头、96孔板型核酸提取试剂盒、自动测序文库构建系统以及测序仪,实现细菌基因组序列测定... 目的建立高通量、半自动化细菌全基因组序列测定体系,满足基于基因组序列的细菌性病原体监测的需求。方法使用自动化液体处理器、可丢弃性机械手枪头、96孔板型核酸提取试剂盒、自动测序文库构建系统以及测序仪,实现细菌基因组序列测定的半自动化。结果机械手枪头与加样孔底的距离、液体混合速度和每次混匀液体的体积比3个因素与样品之间的交叉污染密切相关,机械手枪头与加样孔底的距离为2 mm、液体混合速度为全速的15%、每次混匀液体的体积比为50%时污染率为0。与手工方法相比,高通量半自动化细菌全基因组序列测定体系的工作效率提高了4倍。结论建立的高通量、半自动化细菌基因组序列测定体系,能够满足基于基因组序列的细菌性病原体监测的需求。 展开更多
关键词 自动化液体处理 交叉污染 全基因组序列测定 病原体网络化监测
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Molecular epidemic survey on co-prevalence of scrub typhus and marine typhus in Yuxi city,Yunnan province of China 被引量:18
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作者 zhang Li-juan LI Xue-mei +4 位作者 zhang De-rong zhang jing-shan DI Ying LUAN Ming-chun FU Xiu-ping 《Chinese Medical Journal》 SCIE CAS CSCD 2007年第15期1314-1318,共5页
Background Human rickettsioses are worldwide zoonoses and it is not easy to differentiate them from other infectious diseases because of their atypical manifestation. In recent years the number of patients with fever ... Background Human rickettsioses are worldwide zoonoses and it is not easy to differentiate them from other infectious diseases because of their atypical manifestation. In recent years the number of patients with fever of unknown causes from Hongta District CDC, Yuxi city of Yunnan Province has been increasing significantly in the summer. Diagnosis of scrub typhus was made by local clinicians. In order to ascertain the disease, we undertook a laboratory investigation for such patients from August 18 to 26, 2005. Methods Active surveillance was conducted by Hongta District CDC Yuxi city of Yunnan Province from 2002 to 2004 and basic data were obtained from cases confirmed according to clinical definitions. Average incidences and town-level incidences were calculated during the study periods. Blood samples were analyzed by PCR and serological test. Based on the groEL gene sequences a paired general outer primers (Gro-1 and Gro-2) targeting typhus, spotted fever as well as scrub typhus and two paired inner primers (SF1, SR2 and TF1, TR2) for typhus together with spotted fever and scrub typhus, respectively, were designed to perform a multiplex-nested PCR. Serological assay was carried out by indirect immunofluorescence assay with 7 different rickettsial antigens, i.e., R.mossori, R.sibirica, R.conorii, O.tsutsugamushi, B.quintana, B.henselae and Coxilella burnetii phase Ⅱ Ag. Results Epidemiological surveillance showed that from 2002 to 2004, the average incidences of the scrub typhus or scrub typhus with murine typhus were 222.1/10^5, 204.3/10^5 and 109.6/10^5, respectively. Of 13 blood samples taken during acute stage of illness, 6 showed the amplified products for scrub typhus and the sequenced products showed 100%, 99%, 99%, 99%, 99%, 99% similarity to O.tsutsugamushi Karp but they shared the same deduced amino acid sequences, which indicated 100% identity with the heat shock protein of the O.tsutsugamushi Karp strain. Five yielded PCR products for murine typhus and their corresponding nucleotide sequences exhibited 100%, 100%, 99%, 99% and 99% similarity to R. mossori Wilmington and the analyses of predicted amino acid sequences indicated 100%, 100%, 98%, 98% and 98% identity with the heat shock protein of R. mossoriWilmington strain. Of the 8 PCR positive patients, 3 showed a co-infection of scrub typhus with murine typhus. All the 13 serum samples from febrile patients were positive against O. tsutsugamushi and 8 of them were positive against R. mossori. All of the 8 paired specimens had four-fold elevation of antibody against O. tsutsugamushi, and seroconversion for typhus was demonstrated in 3 paired serum samples. Another finding in the study was that a high seropositive prevalence (76.9%) of Q fever was detected. Concluaion It's confirmed that co-prevalence of scrub typhus with murine typhus are occurring in Yuxi city of Yunnan province, China. Other rickettsial diseases also need to be investigated in these areas. 展开更多
关键词 scrub typhus murine typhus Q fever PREVALENCE Yuxi
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