乙型肝炎患者空腹血清甘胆酸含量测定的临床意义

采用放射免疫分析方法,分别测定了献血员,HBsAg无症状携带者及轻肝活检诊断的各型肝炎患者空腹血清中甘胆酸含量。重症肝炎患者血清中含量最高,急性黄疸型肝炎次之,HBsAg无症状携带者平均值稍高于HBsAg阴性的健康献血员。急性乙型肝炎患者经过治疗,肝功能试验虽已恢复正常,肝组织活检仍可有轻微的损害,血清甘胆酸含量也明显高于健康献血员和HBsAg无症状携带者,提示空腹血清甘胆酸检测,可作为估计肝细胞损害情况的一种辅助方法。     

病毒性肝炎血清前白蛋白与补体C3检测的临床意义

血清前白蛋白(PAB)和补体C3主要由肝脏细胞合成、分泌。为了探讨PAB、C3的含量测定对病毒性肝炎患者的诊断、疗效的观察以及预后判断的临床意义,本文报道45名健康人和186例各型病毒性肝炎患者进行PAB、C3血清含量的检测。材料和方法一、检测对象　1.健康组　健康献血员45名。2.肝炎组　按1995年北京会议的病毒性肝炎防治方案(试行)诊断的肝炎患者186例,各类肝炎的名称和例数参见附表所列。另外对25例重型肝炎患者在治疗过程中进行PAB动态监测。二、方法　美国Beckman公司Array360型全自动蛋白分析仪及试剂,采用速率散射比浊法。结　…     

血清中输血传播病毒检测及临床意义的探讨

目的 研究血清中输血传播病毒（TTV）抗体的检测及其临床意义。方法采用间接ELISA法检测88例患者及20例献血员血清中TTV抗体。结果88例患者中11例血液透析患者、35例乙型肝炎患者、26例丙型肝炎患者、4例庚型肝炎患者和12例非甲-庚型肝炎患者以及20例献血员TTV抗体阳性率分别为27．3％、20．0％、7．8％、0．0％、25．0％与5．0％。结论 在正常人群中TTV健康携带者较为常见；血液透析患者与非甲-庚型肝炎患者是TTV感染的高危人群；乙型及丙型肝炎患者常重叠TTV感染。     

献血员和正常人血清抗HEV-IgG检测结果分析

献血员和正常人血清抗ＨＥＶ－ＩｇＧ检测结果分析凌蓉赵宏王国忠（丹阳市人民医院检验科，江苏丹阳２１２３００）关键词戊型肝炎献血员近年来，国外文献报道戊型肝炎病毒（ＨＥＶ）存在肠道外感染的可能性〔１〕。为此，我们对本地区职业献血员及健康人群进行了抗ＨＥＶ...     

血清载脂蛋白 B 含量测定在慢性肝病中的应用

载脂蛋白 B(Apo B)是低密度脂蛋白的主要载脂蛋白,调节体内胆固醇合成。本文报告81例慢性肝病患者血清 Apo B 含量,以探讨 ApoB 含量与慢性肝病之间的关系。对象和方法一、临床资料(一)正常对照组:74例,男40例,女34例。系本院健康献血员和健康查体者。(二)观察组:肝硬化60例,慢性肝炎21例,临床诊断符合1984年南宁肝炎会议标准。     

输血传播庚型肝炎的研究进展

HGV/GBV C是近年发现的新型肝炎病毒 ,能够引起非甲～非戊型肝炎 ,HGV/GBV C广泛存在于输血后肝炎及献血员中。目前实验室检测HGV/GBV C感染主要依靠RT PCR和ELISA。本文就RT PCR及ELISA检测HGV/GBV C的实验条件和HGV/GBV C在供血员中的感染情况及其引起输血后HGV/GBV C感染率进行介绍 ,并根据目前文献提出了今后输血传播HGV/GBV C的研究方向。     

我国不同肝炎患者及献血员中SENV-D/H感染流行病学调查

目的了解SEN病毒(SENV)D亚型和H亚型在我国不同肝炎患者和献血员中的感染情况。方法采用巢式聚合酶链反应对收集的270例健康献血者和484例各型肝炎患者(包括甲肝、乙肝、丙肝和甲非戊型肝炎患者)进行SENV-D亚型和H亚型DNA检测。结果无偿献血者中SENV-D的感染率为25.9%,SENV-H为24.4%;急性甲肝患者中SENV-D的感染率为14.5%,SENV-H为32.7%;慢性乙肝患者中SENV-D的感染率炎46%,SENV-H为45.2%;在丙肝患者中SENV-D和SENV-H感染率均为65%,在非甲-非戊型肝炎患者中SENV-D的感染率为51.2%,SENV-H为43,9%。结论SENV可能为HCV和HBV具有一样的传播方式,但并不是肝病发生的原因。     

病毒性肝炎患者血中内皮细胞及vWF等变化的初步探讨

本文以１００例血库献血员为正常对照，６０例病毒性肝炎患者为研究对象，在检测了肝炎患者血浆血管性血友病因子相关抗原、补体Ｃ３含量及白细胞吞噬率变化的同时，对循环中内皮细胞的分离和检测方法进行了初步探讨。     

我国不同肝炎患者及献血员中SENV-D/H感染流行病学调查

目的了解SEN病毒(SENV)D亚型和H亚型在我国不同肝炎患者和献血员中的感染情况。方法采用巢式聚合酶链反应对收集的270例健康献血者和484例各型肝炎患者(包括甲肝、乙肝、丙肝和非甲非戊型肝炎患者)进行SENV-D亚型和H亚型DNA检测。结果无偿献血者中SENV-D的感染率为25.9%,SENV-H为24.4%;急性甲肝患者中SENV-D的感染率为14.5%,SENV-H为32.7%;慢性乙肝患者中SENV-D的感染率为46%,SENV-H为45.2%;在丙肝患者中SENV-D和SENV-H感染率均为65%;在非甲-非戊型肝炎患者中SENV-D的感染率为51.2%,SENV-H为43.9%。结论SENV可能与HCV和HBV具有一样的传播方式,但并不是肝病发生的原因。     

TT病毒核酸的检测

目的初步测定献血员、慢性乙型肝炎、慢性丙型肝炎及慢性非甲～戊型，非庚型肝炎患者中ＴＴ病毒的感染情况，分析不同ＴＴ病毒感染者血清中ＴＴ病毒开放读码框架２区部分基因序列。方法以套式聚合酶链反应测定ＴＴ病毒核酸，取献血员（ＴＸ２）、慢性乙型肝炎（ＴＸ３）、慢性丙型肝炎（ＴＸ４）及慢性非甲～戊型，非庚型肝炎患者（ＴＸ１）各１例ＴＴ病毒ＰＣＲ阳性产物，以双脱氧核苷酸链终止法测定核苷酸序列。结果检测２０例献血员、２９例慢性乙型肝炎、３１例慢性丙型肝炎及３２份慢性非甲～戊型，非庚型肝炎标本，ＴＴ病毒核酸阳性者分别占５．０％（１／２０）、６．９％（２／２９）、２９．０％（９／３１）和３４．４％（１１／３２）。与Ｎ２２克隆相比，ＴＸ１、ＴＸ２、ＴＸ３与ＴＸ４在核苷酸水平的同源性分别为９５．４１％、９８．４７％、９７．４５％和９７．９６％。结论本组献血员、慢性乙型肝炎、慢性丙型肝炎及慢性非甲～戊型，非庚型肝炎患者中均存在ＴＴ病毒感染者。从本组不同ＴＴ病毒感染者血清中分离出４株ＴＴ病毒序列，其开放读码框架２区部分基因序列高度保守     

中国人群肥厚型心肌病患者心肌肌钙蛋白基因突变研究

目的:初步探讨中国人群肥厚型心肌病患者心肌肌钙蛋白T基因突变情况,为该病的早期防治和康复介入奠定理论基础。方法:提取58例肥厚型心肌病(hypertrophiccardiomyopathy,HCM)患者外周血白细胞基因组DNA,聚合酶链反应(PCR)扩增心肌肌钙蛋白T(cTnT)基因8、9、11号外显子,产物做单链构象多态性(SSCP)分析,出现异常条带者直接测序验证。结果:58例临床确诊的HCM患者cTnT基因8,9,11号外显子,除160位密码子存在ATC和ATT多态性外,未发现基因位点的异常。结论:与国外报道的肥厚型心肌病约20%是由肌钙蛋白T基因突变引起相比较,中国人群肥厚型心肌病患者基因突变可能存在不同的特点。     

深圳献血人群隐匿性乙型肝炎病毒全基因序列系统进化树分析

目的了解深圳献血人群隐匿性乙型肝炎B和C基因型及亚型的分布规律,研究不同毒株全基因序列进化关系。方法对30例HBsAg(-)/HBV DNA(+)标本进行基本核心启动子/前C区(BCP/PC,295 bp)、HBV全基因(3 162 bp)巢式PCR扩增,PCR产物克隆后测序。2者均为阳性的5份标本合成3 215 bp长的全基因序列,与GenBank中已发表的HBV A～H基因型23株的全序列进行系统进化树分析确定基因型,将所属基因型B和C亚型再作系统进化树分析,以确定基因亚型。结果获得5例全基因序列,4例为C型,1例为B型。分属于C2,C2,C2,C1和B2亚型。3例C2亚型与日本、马来西亚基因亚型的进化距离最近,C1株与马来西亚和泰国2例携带者的病毒株进化距离最近。B2株与印度尼西亚华裔基因亚型的病毒株进化距离最近。结论深圳献血人群隐匿性乙型肝炎感染病毒基因亚型有C2,C1,B2,以C2亚型为主。     

Detection of TT virus in hemodialysis patients]

Recently a newly discovered DNA virus, transfusion transmitted virus (TTV), was introduced as a cause of post-transfusion hepatitis. We studied the frequency of TTV viremia in 60 hemodialysis patients in Yamaguchi, Japan. TTV DNA was detected by heminested PCR, using primers described by Okamoto et al. TTV DNA was detected in 18 patients (30%). There was no differences in clinical characteristics, including age, gender, history of blood transfusion, and double infection of other hepatitis viruses, between TTV DNA positive patients and negative patients. Also the frequency of TT viremia was not associated with the duration of hemodialysis. These results suggest that the routes of TTV infection may be different from those of infection by HBV, HCV, or HGV.     

The use of polymerase chain reaction to detect septicemia in critically ill patients.

OBJECTIVE: To describe the use of bacterial DNA amplification of conserved bacterial 16S ribosomal DNA nucleotide sequences by polymerase chain reaction (PCR) to detect the presence of septicemia in critically ill septic patients. DESIGN: Case series of blood samples from septic patients comparing the PCR results with conventional blood culture results. SETTING: A general intensive care unit in a tertiary referral hospital. PATIENTS: Two sets of samples (n = 101 and n = 55) from patients diagnosed as clinically septic and requiring blood cultures. They were classified by internationally accepted criteria into systemic inflammatory response syndrome, severe sepsis, and septic shock groups. INTERVENTIONS: Blood samples taken in a sterile fashion concurrently for blood culture, and PCR of the bacterial 16S ribosomal RNA gene in leukocytes and plasma. Two different DNA extraction techniques for PCR were tried sequentially. MEASUREMENTS AND MAIN RESULTS: Blood culture and PCR positivity were measured in relation to the clinical classification of severity of sepsis. Using the initial extraction method (n = 101), ten patients were positive by both PCR and blood culture, eight patients were PCR positive and blood culture negative, and seven patients were blood culture positive and PCR negative. From the clinical criteria, PCR detected at least six true positives that had been missed on blood culture and missed four true Gram-positive bacteremias. When the initial code was broken, this deficiency was rectified using the improved extraction technique (n = 55), in which ten patients were positive by PCR and blood culture, 29 patients were PCR positive and blood culture negative, and two patients were PCR negative and PCR positive. CONCLUSIONS: We conclude that the use of PCR (for the 16S ribosomal DNA in the plasma) was significantly more sensitive than the use of conventional blood culturing techniques for the detection of bacteremia in seriously ill patients. This could prove to be a valuable adjunct to conventional blood cultures.     

Duffy血型基因定型及其用于HDN产前诊断的探讨

目的　探讨用分子生物学方法检测血液Duffy血型基因型及该方法应用于HDN产前检查的可行性。方法　采用聚合酶链反应 限制性酶切片短长度多态性 (PCR RFLP)方法检测Duffy基因型。结果　用本研究采用的方法检出了 90例中国辽宁地区汉族血液样品Duffy血型的 3种基因型 ;检测出丈夫为Fy(a+b - )型、本人为Fy(a -b +)型的孕妇羊水样品 1例 ,其Duffy基因型为FYAFYB。结论　本方法可准确检测血液及羊水样品的Duffy基因型 ,可用于Duffy血型不合引起的HDN的产前诊断     

Real-time PCR for detection of Trypanosoma brucei in human blood samples

We have developed a real-time PCR assay for detection of Trypanosoma brucei DNA in human blood samples. The PCR was conducted with newly designed primers targeting the 177-bp repeat satellite DNA in T. brucei and with Sybr Green to monitor the amplicon accumulation. DNA purification using Chelex 100 resin was performed on blood samples collected on Whatman FTA cards and was shown to be a simple and quantitative method as revealed by real-time PCR. The detection limit of the assay was 100 trypanosomes per mL blood, corresponding to an analytical sensitivity of 0.1 genome equivalents. Trypanosome DNA was detected in all blood samples from sleeping sickness patients and, furthermore, the identity of the amplicon was confirmed in all assays by dissociation analysis. Although template DNA from blood samples was amplified with significantly lower efficiency than genomic DNA, similar efficiency between all assays ensured quantitative results. No amplicon product was obtained with samples from uninfected individuals. The results indicate that the real-time PCR assay described is a rapid and sensitive method suitable for the detection of T. brucei in human blood samples in routine clinical laboratory practice.     

Carboxyl-terminal sequence variation of latent membrane protein 1 gene in Epstein-Barr virus-associated gastric carcinomas from Eastern China and Japan

OBJECTIVES: To elucidate variations of latent membrane protein 1 (LMP1) in Epstein-Barr virus (EBV)-associated gastric carcinoma (EBVaGC) and explore the LMP1 variations of neighboring countries, China and Japan. METHODS: In 12 and 8 EBVaGCs from eastern China and Japan, respectively, the C-termini of LMP1 were analyzed using PCR and sequencing. The sequences were compared with previously published strains and were characterized on a phylogenetic tree. The difference between Chinese and Japanese isolates was characterized. RESULTS: Ten of 12 Chinese GC isolates (83.3%) and all of the 8 (100%) Japanese GC isolates belonged to the China 1 strain. Also, B95-8 type isolates were found in 2 of 12 Chinese GC. In the 18 China 1 type isolates, additional mutations outside the signature sequence changes were found. All Japanese isolates (100%) had two or more additional mutations, whereas only 5 of 10 (50%) Chinese isolates had two or more additional mutations. The difference was statistically significant (p = 0.0359). CONCLUSIONS: China 1 is the dominant strain in GC from eastern China and Japan. The similarity to that of nasopharyngeal carcinoma (NPC) from China supports the view that China 1 strain represents a geographic-associated polymorphism rather than an NPC-associated polymorphism. Japanese isolates show more mutations than Chinese isolates, suggesting a geographic difference between Chinese and Japanese isolates in GC.     

家族性帕金森病患者parkin基因缺失突变的初步研究

目的探讨中国家族性帕金森病( parkinson's disease, PD)患者 parkin基因第 3～ 7外显子是否存在缺失突变,及其与该病临床特点的关系. 方法采集 6例无血缘相关的家族性 PD患者外周血液,提取 DNA,通过 PCR扩增、琼脂糖凝胶电泳鉴定 parkin基因第 3～ 7外显子缺失突变,并结合临床资料分析. 结果 6例无血缘相关的家族性 PD患者中,发现 1例有第 5外显子缺失,其遗传模式呈常染色体隐性遗传,起病年龄 60岁,临床表现为震颤、僵直和运动迟缓,但无异动症.第 3, 4, 6, 7外显子未发现缺失突变. 结论中国家族性 PD患者中存在 parkin基因第 5外显子缺失突变改变.     

聚合酶链反应扩增伤寒患者粒细胞中伤寒沙门菌DNA

目的建立适用于临床应用的血中伤寒沙门菌（Ｓ．ｔｙｐｈｉ）ＤＮＡ扩增的方法。方法分离患者粒细胞，采用裂解法制备模板ＤＮＡ，单管套式聚合酶链反应扩增Ｓ．ｔｙｐｈｉ鞭毛基因。结果６例血培养阴性而临床拟诊伤寒及２３例由血培养确诊的伤寒患者粒细胞均获得了３４３ｂｐ的扩增产物，其他３４例发热待查患者粒细胞无扩增产物出现。连续１１０稀释Ｓ．ｔｙｐｈｉＤＮＡ，最低可检测到４０ｆｇＤＮＡ（约１０个菌）。扩增产物的ＤＮＡ序列测定结果也证实了３４３ｂｐ的扩增产物是Ｓ．ｔｙｐｈｉ鞭毛基因。结论该方法是一项简便、省时、灵敏和特异的实验室诊断伤寒的新技术，尤其对培养阴性的伤寒患者可提供早期、快速的实验室诊断，套式扩增反应在同一反应管中完成，能有效避免标本间的污染。     

Improvement in the laboratory recognition of lyme borreliosis with the combination of culture and PCR methods.

BACKGROUND: Lyme disease is a multisystem, multistage infection caused by three genospecies of the Borrelia burgdorferi sensu lato species. The diagnosis of Lyme disease is based on a history of tick-bite, physical examination, and serological tests. In the seronegative patients with Lyme borreliosis symptoms, additional testing should be introduced. METHODS: The study group was composed of 240 hospitalized patients presented with various clinical symptoms suggesting Lyme borreliosis: 221 of the patients with neurological abnormalities and 19 with oligoarticular arthritis. Citrated blood and serum samples were collected from the patients for culture and serological examination, respectively. Moreover, 173 cerebrospinal and 6 synovial fluid samples were tested. New oligonucleotide primers based on B. burgdorferi sensu lato 16SrRNA gene sequences were designed for the detection of the bacteria in blood, cerebrospinal, and synovial fluid specimens with PCR. Levels of specific antibodies were measured in serum, cerebrospinal fluid and synovial fluid samples using ELISA and Western blot. B. burgdorferi spirochetes from blood, cerebrospinal fluid, and synovial fluid samples were cultured in cell line. Extracted and purified B. burgdorferi DNA was identified by PCR with new oligonucleotide primers. Then three genospecies were identified by PCR amplification with other primer sets specific for 16S rDNA and/or by the restriction fragment length polymorphism of 23S(rrl)-5S(rrf). RESULTS: Bacterial DNA were found in samples from 32 patients, including 28 patients with neuroborreliosis and 4 with Lyme arthritis. B. burgdorferi-specific IgM and/or IgG serum antibodies were detected in 14 of these patients. Fourteen strains of Borrelia garini, 4 strains of Borrelia afzelii and 1 strain of B. burgdorferi sensu stricto were identified by PCR. Genospecies were not recognized in 13 specimens. CONCLUSIONS: The procedure can be a rapid and sensitive diagnostic method for the detection of etiological agents in clinical materials derived from patients with the clinical symptoms of Lyme borreliosis. It can be utilized for both basic research as well as routine laboratory diagnosis.