肝癌实验室诊断研究进展

原发性肝癌是目前世界上致死率最高的恶性肿瘤之一,在全世界范围内的发病率有逐年上升的趋势。肝癌的诊断特别是早期诊断,对于提高患者生存率至关重要。我们简要综述了国内外肝癌实验室诊断中的常用标志物以及检测方法的进展,并展望了其发展前景。     

淋球菌感染实验室诊断的实用技术

淋病奈瑟菌 (N eisseria gonorrhoeae)是淋病的病原体 ,该菌也称淋病双球菌 (diplococcus gonorrhoea)或淋球菌(gonoccus)。淋球菌的分离培养检测法仍是诊断淋病的金标准试验技术 ,基因探针技术有可能比分离培养法更敏感和特异而成为另一种可行的诊断方法 [1 ] 。在设计淋病的诊断技术时 ,实验技术人员要考虑所在单位的实际、社区淋病患病率以及试验方法的成本效益等 ,试验效果 (testperformance)是选择淋病诊断试验的关键指标。当社区淋病患病率低时 (如产前检查的妇女 ) ,敏感度与特异度均高的试验方法的阳性预测值 (PPV)也低 ;类似试验…     

细小病毒B19感染实验室诊断技术的研究进展

人类细小病毒感染,临床可有多种表现,尤其可导致胎儿早产、流产、死胎、贫血、发育迟缓及形成积水型胎儿。本文拟从电镜检查、ＥＬＩＳＡ、固相放射免疫技术（ＲＩＡ）、分子杂交及多聚酶链反应（ＰＣＲ）等几个方面对该病毒感染实验室诊断的国外最新研究进展作一介绍。     

衣原体实验室诊断技术研究进展及其前景

对衣原体的分子生物学特性和各种实验技术及其研究进展作了简要综述。     

人兽共患细菌病的流行现状及其病原耐药性的应对策略

人兽共患病是由同一病原引起、在人和脊椎动物之间自然传播的疾病,已成为全球共同面临并亟待解决的重大问题。当前人兽共患传染病在人类新发和现有传染病中占比达60%,其中布鲁氏菌(Brucella)、牛分枝杆菌(Mycobacterium bovis)、炭疽芽胞杆菌(Bacillus anthracis)、猪链球菌2型(Streptococcus suis serotype 2, SS2)、沙门氏菌(Salmonella)及肺炎克雷伯氏菌(Klebsiella pneumoniae, KP)等引起的人兽共患细菌病严重威胁畜牧业健康发展、公共卫生安全和国家生物安全,其防控形势日益严峻。本文就上述6种重要人兽共患细菌病的流行现状及其相关耐药机制和抗菌新策略进行综述,为科学防控人兽共患细菌病提供参考。     

布鲁氏菌病关节炎的致病机制研究进展

布鲁氏菌病(布病)是由布鲁氏菌感染引起的一种人畜共患传染病.动物与人感染布鲁氏菌都能表现累及关节相关的症状,即布病关节炎.典型的动物病理模型表明,布鲁氏菌关节炎发病时关节腔周围组织有大量中性粒细胞、单核细胞和破骨细胞的浸润并伴有明显的骨破坏.除了布鲁氏菌对骨关节细胞的直接作用外,布鲁氏菌和骨组织细胞、免疫细胞还通过细胞...     

检测OMP28抗体不能有效诊断羊布鲁氏菌病

【目的】探索布鲁氏菌外膜蛋白OMP28作为羊布鲁氏菌病特异性检测方法的可行性。【方法】体外表达和纯化OMP28蛋白,建立并优化以OMP28重组蛋白为包被抗原的布鲁氏菌病ELISA诊断方法。以3个不同种属的4株布鲁氏菌(羊种布鲁氏菌16M和M28,猪种布鲁氏菌S1330,牛种布鲁氏菌2308)分别感染山羊和绵羊至42周,期间每隔2周收集血清,分别用布鲁氏菌LPS包被的ELISA和OMP28 ELISA方法对不同阶段的分离血清进行检测,比较2种不同ELISA对4株布鲁氏菌感染的山羊和绵羊的检测敏感性。【结果】4株布鲁氏菌感染的山羊和绵羊均产生高水平针对LPS的抗体,但是仅有B. melitensis 16M和B. melitensis M28感染的绵羊与B. melitensis 16M和B. abortus 2308感染的山羊可产生针对OMP28的抗体。【结论】基于OMP28的间接ELISA具有细菌属特异性和宿主动物品种特异性,通过检测OMP28抗体不能有效诊断羊布鲁氏菌病。     

布鲁氏菌病荧光偏振抗体检测方法的建立

【目的】布鲁氏菌病(Brucellosis)简称布病,是由布鲁氏菌引起的以感染家畜为主的人畜共患传染病,造成严重的公共卫生问题。目前全世界范围内消除该病的主要方法是扑杀与免疫相结合,所以建立快速准确的诊断方法对防治和清除布病非常必要。本文建立布鲁氏菌病荧光偏振(FPA)抗体检测方法,为布鲁氏菌病(布病)的快速高效诊断提供技术手段。【方法】提纯猪种布鲁氏菌S2株脂多糖O链(OPS),经异硫氰酸荧光素(FITC)标记后作为诊断抗原。通过对样品稀释液、抗原稀释度、反应时间等条件的优化,初步建立了布鲁氏菌荧光偏振诊断方法。用该方法对148份布病阳性血清(其中牛血清70份,羊血清78份)和155份布病阴性血清(其中牛血清82份,羊血清73份)进行检测,确定其敏感性和特异性。按确定的技术参数,制备3批布鲁氏菌FPA抗体检测试剂盒,使用质控阴、阳性血清分别评价试剂盒的批内和批间重复性。用400份临床样本比较本研究开发试剂盒与商品化进口FPA试剂盒的符合率。【结果】使用0.5%蔗糖磷酸缓冲液作为血清样品稀释液;标记抗原的使用浓度为90μg/m L;最佳反应时间为3-5 min。本检测方法的判定标准为:δm P值20时为阴性,δm P值≥20时为阳性。按上述条件建立的FPA检测148份布病阳性血清和155份布病阴性血清,结果敏感性为98.6%,特异性为98.7%。对400份临床样本的比对检测显示,研究建立的FPA方法与进口商品化试剂盒的总符合率为94.0%。【结论】研究建立的布鲁氏菌PFA抗体检测方法具有良好的特异性和敏感性,可作为一种重要的布病诊断快速诊断方法。     

诺如病毒的致病机制及诊断

诺如病毒(Norovirus, NoV)是引起全人群急性胃肠炎暴发和流行的主要病原体,也是引起食源性疾病的最常见非细菌性病原体。由于缺少有效的小型动物及培养细胞研究模型,目前对NoV感染的致病机制尚不清楚。NoV实验室诊断技术发展成熟,尤其是近年来新技术的应用将进一步推动其诊断水平的提高,满足公共卫生和临床诊疗的新需求。     

珊瑚病原菌株XSBZ03和XSBZ14双重PCR检测方法的建立

【目的】建立珊瑚病原菌株XSBZ03和XSBZ14的双重PCR检测方法。【方法】以XSBZ03和XSBZ14的特异靶序列为对象,开展引物设计和双重PCR检测方法的构建,并确定该双重PCR方法的特异性、敏感性及可靠性。【结果】该检测方法可特异识别菌株XSBZ03和XSBZ14,对XSBZ03和XSBZ14基因组DNA样品的检测极限分别为1.7pg/μL和2.0pg/μL;对XSBZ03和XSBZ14在海水样品中的检测极限分别为6×10～3 CFU/mL和8×10～3 CFU/mL。【结论】该方法具有特异性强、灵敏度高等优点,可对由菌株XSBZ03和XSBZ14引起的珊瑚疾病进行准确快速的诊断,为今后开展珊瑚疾病防控和无特定病原的珊瑚移植提供了可靠手段。     

农村医师专业实验诊断学实践教学改革过程中的体会及探讨

目的：探讨传统教学法和多媒体教学法两种教学方法在农村医师专科实验诊断学实践教学中的应用及比较。方法：在实验诊断实验教学过程中,通过对农村医师专科2011-9班采用传统教学法,对农村医师专科2011-7班采用多媒体教学法,并应用统计学方法对两个班的实验考试平均成绩和及格率进行比较和分析。结果：农村医师专业2011-9班的平均成绩为75.65分,2011-7班平均成绩为84.44分。对平均成绩进行F检验,F=1.12,P〉0.10,方差齐,故可用两样本t检验,t=5.36,P〈0.01;农村医师专科2011-9班的及格率为68.09%,农村医师专科2011-7班的及格率为86.37%,X2=4.49,P〈0.05。采用多媒体教学法的农村医师专科2011-7班的平均成绩和及格率明显高于采用传统教学法的农村医师专科2011-9班。结论：在农村医师专科实验诊断学实验教学中,多媒体教学法跟传统教学法比具有明显的优势。     

Investigating zoonotic infection barriers to ape Plasmodium parasites using faecal DNA analysis

African apes are endemically infected with numerous Plasmodium spp. including close relatives of human Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale, and Plasmodium malariae. Although these ape parasites are not believed to pose a zoonotic threat, their ability to colonise humans has not been fully explored. In particular, it remains unknown whether ape parasites are able to initiate exo-erythrocytic replication in human hepatocytes following the bite of an infective mosquito. Since animal studies have shown that liver stage infection can result in the excretion of parasite nucleic acids into the bile, we screened faecal samples from 504 rural Cameroonians for Plasmodium DNA. Using pan-Laverania as well as P. malariae- and P. vivax-specific primer sets, we amplified human P. falciparum (n?=?14), P. malariae (n?=?1), and P. ovale wallikeri (n?=?1) mitochondrial sequences from faecal DNA of 15 individuals. However, despite using an intensified PCR screening approach we failed to detect ape Laverania, ape P. vivax or ape P. malariae parasites in these same subjects. One faecal sample from a hunter-gatherer contained a sequence closely related to the porcupine parasite Plasmodium atheruri. Since this same faecal sample also contained porcupine mitochondrial DNA, but a matching blood sample was Plasmodium-negative, it is likely that this hunter-gatherer consumed Plasmodium-infected bushmeat. Faecal Plasmodium detection was not secondary to intestinal bleeding and/or infection with gastrointestinal parasites, but indicative of blood parasitaemia. Quantitative PCR identified 26-fold more parasite DNA in the blood of faecal Plasmodium-positive than faecal Plasmodium-negative individuals (P?=?0.01). However, among blood-positive individuals only 10% - 20% had detectable Plasmodium sequences in their stool. Thus, faecal screening of rural Cameroonians failed to uncover abortive ape Plasmodium infections, but detected infection with human parasites, albeit with reduced sensitivity compared with blood analysis.     

禽流感病毒实验检测研究进展

禽流感病毒(avian influenza viruses,AIV)给人类健康已带来严重威胁,而实验室快速、准确的诊断技术对禽流感的预防、控制及应急反应决策起着极其关键的作用,这使其成为了研究热点并取得了巨大进步。就禽流感的实验检测技术研究进展从病毒分离、免疫学诊断及分子诊断3个方面加以综述。     

Blastocystis subtype 5: Predominant subtype on pig farms,Thailand

Blastocystis is a unicellular protist most commonly detected in humans and a variety of animals. The predominant mode of its transmission is the fecal–oral route, but its zoonotic potential is not completely understood. The objective of this study was to determine the presence and genetic diversity of Blastocystis on pig farms in Nakhon Pathom Province, Central Thailand. A total of 154 human and 90 pig stool samples were collected and analyzed. Nested PCR detected Blastocystis in 35.55% of the pig samples and 6.49% of the human samples. Subtyping based on regions of the small-subunit ribosomal RNA (SSU rRNA) gene identified three Blastocystis subtypes in pigs and humans: ST1, ST3, and ST5. Blastocystis ST5 was the predominant subtype, followed by ST1 and then ST3. All the sequences from the Blastocystis-positive samples from both pigs and humans were closely related. This study reveals a possibility of low host specificity of Blastocystis STs (ST1, ST3 and ST5) on pig farms in Thailand. We tentatively suggest that close contact with or exposure to pig stools may be a significant source of Blastocystis detected in pig handlers. Further studies are required to confirm the zoonotic transmission of this organism in Thailand, because pigs may play an important role in the transmission of Blastocystis.     

The zoonotic,fish-borne liver flukes Clonorchis sinensis,Opisthorchis felineus and Opisthorchis viverrini

Clonorchis sinensis, Opisthorchis felineus and Opisthorchis viverrini are the three most important liver flukes involved in human health, infecting more than 45 million people worldwide. Both C. sinensis and O. viverrini, and possibly O. felineus, can induce human cholangiocarcinoma as well as inducing other hepatobiliary pathology. Although the life cycles of all three species are similar, only that of O. felineus in Europe remains predominantly zoonotic, while O. felineus in Asia and C. sinensis have a stronger mixture of zoonotic and anthroponotic components in their life cycles. Opisthorchis viverrini from the Mekong area of southeastern Asia is predominantly anthroponotic. Here we discuss the comparative epidemiology of these three taxa comparing in detail the use of first, second and final animal hosts, and consider the potential role of humans in spreading these pathogens. In addition we discuss the genetic structure of all three species in relation to potentially cryptic species complexes.     

A natural zoonotic giardiasis: Infection of a child via Giardia cysts in pet chinchilla droppings

Here, we report a case of direct zoonotic transmission of giardiasis between a pet chinchilla and a human. Microscopic and molecular examinations of stool samples from a child and samples of chinchilla droppings revealed cysts/DNA of Giardia intestinalis. The transmission from the chinchilla to the child has been confirmed as coprophagous after the 1-year-old toddler ingested pet chinchilla droppings. Molecular analysis of the gdh gene from both hosts classified the G. intestinalis cysts into the assemblage B genetic group, which has been previously shown to be characteristic of both human and chinchilla giardiasis. Both Giardia sub-assemblages BIII and BIV were present in the chinchilla droppings, whereas only the sub-assemblage BIV was isolated from the child's stool sample. To the best of our knowledge, this is the first report of a true zoonotic transmission of giardiasis, supporting the zoonotic potential of assemblage B.     

非幽门螺杆菌：一类人兽共患病病原菌

过去的几十年,非幽门螺旋杆菌的研究迅速发展,他们不仅能感染和人类密切相关的动物,在人类的胃肠、肝胆及其他系统的疾病中也起着重要的作用,是一类人兽共患病病原菌。