登革病毒的分子生物学检测技术研究进展

登革病毒是一种蚊媒急性传染病的病原体，属于黄病毒属，所致疾病主要有登革热（dengue fever／DF）、登革出血热（Dengue hemorrhagic fever／DHIF）和登革休克综合征（Dengue shock syndrome／DSS）。在临床上，登革热主要表现为高热、头痛、肌肉关节痛等，同时伴有白细胞减少；登革出血热和登革休克综合征是登革热的严重类型，多发生于登革热地方流行区的当地居民，以高热、出血、休克和高病死率为特征。     

登革病毒的分子生物学检测技术研究进展

登革病毒是一种蚊媒急性传染病的病原体,属于黄病毒属,所致疾病主要有登革热(dengue fever/DF)、登革出血热(Dengue hemorrhagic fever/DHF)和登革休克综合征(Dengue shock syndrome/DSS)。在临床上,登革热主要表现为高热、头痛、肌肉关节痛等,同时伴有白细胞减少;登革出血热和     

登革病毒动物模型的研究进展

登革病毒(dengue virus)属于黄病毒属，是一种以蚊虫为传播媒介的RNA病毒。根据E蛋白抗原性的不同，登革病毒被分为四个血清型(DEN-1，2，3，4)，它威胁着数以亿计的、居住在热带和亚热带地区的人们的健康。引起登革热(dengue fever，DF)和登革出血热(dengue hemorrhagic fever，DHF)，前者是一种轻型的发热性疾病，而后者则是一种威胁患者生命的综合症，死亡率较高。     

登革病毒实验室诊断研究进展

登革病毒（Dengue virus,DV）是黄病毒科,黄病毒成员,通过蚊子传播,伊蚊是主要传播媒介。病毒感染后可发生登革热、登革出血热、登革休克综合征。广泛流行于热带和亚热带地区,近几十年来,登革热病例急剧增长,据WHO估计,目前全球2／5的人口受到登革病毒感染的威胁,每年约有0．5亿到1亿人感染登革病毒,24000人死亡。     

宿主异常免疫反应与登革病毒致病的关系

登革病毒感染在全世界范围内流行的越来越频繁，爆发日益增加，发生登革出血热及登革休克综合症（DHF／DSS）的病例数也呈现增加的趋势。每年，全球范围内有将近一亿人感染登革热，其中大约有50万例登革出血热。目前，登革病毒引起的DHF／DSS的机制尚未完全明了，本文主要就宿主免疫功能异常与登革病毒致病的关系的研究进展作一综述。     

登革病毒感染树突状细胞的分子机制研究进展

1 登革病毒的流行病学及生物学特征 1.1 流行分布特点 世界分布:登革病毒(DENV)由埃及伊蚊和白纹伊蚊传播,在热带和亚热带地区广泛分布,随着城市化进程逐步向城市扩散.登革病毒感染能引起登革热(DF)、登革出血热(DHF)和登革休克综合征(DSS).登革热流行的特点是出现突然、来势猛、传播快.据WHO估计,最近50年全世界感染率增加了30倍,每年大约有1亿人感染,50万人患登革出血热、登革休克综合症,其中2.5万人死亡,并且患者多数是15岁以下的儿童[1].尽管登革病毒感染严重威胁人类的健康,但目前还没有有效的治疗方法.     

登革病毒疫苗研究进展

登革病毒(Dengue virus,DENV)是登革热、登革出血热和登革休克综合征(DHF、DSS)的病原体,伊蚊为主要传播媒介,广泛流行于全球热带及亚热带的100多个国家和地区,超过25亿人受到登革病毒感染的危胁[1].据估计每年有5000万～1亿登革热患者,25～50万登革出血热患者,登革出血热与登革休克综合征的病死率高达10%～15%[2].近年来随着全球气候变暖、旅游和交通事业发展,伊蚊的分布范围不断扩大,登革热已成为世界上分布最广、发病人数最多、危害最大的虫媒传染病之一.     

登革病毒基因组核苷酸变异与毒力的关系

登革病毒是一种流行于热带、亚热带地区的蚊媒病毒，其引起的登革热、登革出血热／登革休克综合征（DSS／DHF）已成为严重威胁人类健康的公共卫生问题。登革病毒缺乏精确的复制校正系统．在传播过程中核苷酸的变化会导致病毒毒力的变化，目前其致病与免疫机制尚未明了，缺乏有效的疫苗。本文就登革病毒基因变异与其毒力关系方面的研究进行综述。     

墨西哥登革病毒感染者HLA-DR抗原频率研究：DR4可能是DHF的遗传抵抗因素

登革病毒 (DV)属黄病毒属 ,它以蚊虫为媒介 ,引起登革热 (DF)和登革出血热 /休克综合症 (DHF/DSS)。全世界每年DF病例超过 1亿 ,其中 5～ 10 %可发展为死亡率较高的DHF。登革病毒感染已成为热带和亚热带地区严重的公共卫生问题。作者以墨西哥Morelos州 1997～ 1999期间的 34名DHF患者和 4 7名DF患者为研究对象。对患者血液标本进行DNA抽提 ,通过PCR SSO反向斑点杂交对HLA分型 ,再通过PCR SSOP对HLA DRB1 0 4分型并克隆测序。期望通过基因频率的比较找出HLA DRB1等位基因和DHF/DSS发生的联系。结果 :DRB1 0 4等位基…     

登革病毒感染小鼠细胞免疫功能的变化

登革病毒感染小鼠细胞免疫功能的变化郭庆福詹轶群周洁李胜华梁小兵代庆华为探讨细胞免疫在登革热和登革出血热发病中的作用，用Ⅱ型登革病毒的鼠脑毒种（约１０６ｃｐｕ）静脉感染成年ＢＡＬＢ／ｃ小鼠为模型，并以注射同剂量正常鼠脑的小鼠为对照，分成不同时间组，每组...     

广州市2006年Ⅰ型登革病毒流行株E基因序列分析

目的测定广州市2006年Ⅰ型登革病毒流行株的E基因序列并进行分析。方法收集广州市2006年登革热患者急性期血清,用C6／36细胞培养分离登革病毒,RT-PCR法扩增全长E基因,测定序列并绘制系统发生树,进行生物信息学分析。结果59份标本中38份病毒分离培养阳性,获得广州市2006年Ⅰ型登革病毒流行株GZ2006／1707的E基因序列,其同源性与东南亚缅甸、泰国、柬埔寨等地的流行株接近,但与广州市2002年Ⅰ型登革病毒流行株GZ2002／281较远。结论广州市2006年流行的登革病毒属输入性,但与2002年流行的登革病毒有不同的输入源。     

KIR copy number variations in dengue-infected patients from northeastern Thailand

Killer immunoglobulin-like receptors (KIRs) are a family of receptors expressed on Natural killer (NK) cells. The extensive polymorphism of KIR is involved in the immune responses of NK cells and influences dengue infections. We investigated the diversity of KIR copy numbers in dengue-infected patients from northeastern Thailand. Copy numbers of KIRs were determined by quantitative polymerase chain reaction in 137 dengue-infected patients, comprising 63 dengue fever (DF) and 74 dengue hemorrhagic fever (DHF). The distribution of KIRs was observed to be between 0 and 4 copies. The KIR AA genotype with heterozygous KIR2DS4D/WT was the most common in dengue patients, 25.4% DF and 23% DHF. Forty KIR profiles were determined in dengue patients, including 31 usual, 6 expanded, and 3 contracted profiles. Investigation of KIR copy number and dengue severity indicated that two copies of KIR2DL3 combined with HLA-C1C1 associated with an increased risk of DHF (OR 2.32, 95% CI 1.159–4.624, P = 0.016), whereas one copy of KIR2DL2 and KIR2DL3 together with HLA-C1C1 associated with a reduced risk of DHF (OR 0.17, 95% CI 0.058–0.482, P < 0.001). The outcomes of this study will contribute to the understanding of KIR complexity and innate immune responses in dengue infections.     

Central nervous system and muscle involvement in dengue patients: A study from a tertiary care center

BackgroundNeurological involvement in dengue virus (DENV) infection is being increasingly reported. There is paucity of studies evaluating the relative frequency of central nervous system (CNS) and muscle involvement in dengue.ObjectivesTo evaluate the frequency and prognosis of neurological and muscle involvement in dengue, and correlate these with dengue subtypes.Study designConsecutive dengue patients were included, and their clinical features, laboratory investigations and cerebrospinal fluid (CSF) findings were recorded. Cranial MRI was done in unconscious patients and electromyography and nerve conduction study in patients with flaccid weakness. Patients were categorized into encephalopathy, encephalitis, immune mediated and dengue associated muscle dysfunction (DAMD). Outcome at 1 month and its predictors were evaluated.Results116 patients aged 5–70 years were included; 82 had dengue fever (DF), 18 had dengue hemorrhagic fever (DHF), and 16 had dengue shock syndrome (DSS). Neurological manifestations were present in 92 (79%); encephalopathy in 17 (15%), encephalitis in 22 (19%), transverse myelitis in 1 (1%) and DAMD in 52 (45%) patients. Central nervous system (CNS) involvement was commoner in DHF/DSS compared to DF (44% vs 26%). 10 patients with CNS involvement died versus 1 with DAMD. The patients in the CNS group had more frequent hypotension, renal dysfunction and respiratory failure compared to the DAMD group, and had worse outcome. DENV2 and DENV3 were the commonest serotypes, but serotypes did not differ between CNS and DAMD groups.ConclusionsDAMD is commoner than CNS involvement in dengue. CNS involvement however, is associated with more serious illness and predicts poorer outcome.     

Serum cytokine/chemokine profiles in patients with dengue fever (DF) and dengue hemorrhagic fever (FHD) by using protein array

BackgroundDENV infection can induce different clinical manifestations varying from mild forms to dengue fever (DF) or the severe hemorrhagic fever (DHF). Several factors are involved in the progression from DF to DHF. No marker is available to predict this progression. Such biomarker could allow a suitable medical care at the beginning of the infection, improving patient prognosis.ObjectivesThe aim of this study was to compare the serum expression levels of acute phase proteins in a well-established cohort of dengue fever (DF) and dengue hemorrhagic fever (DHF) patients, in order to individuate a prognostic marker of diseases severity.Study designThe serum levels of 36 cytokines, chemokines and acute phase proteins were determined in DF and DHF patients and compared to healthy volunteers using a multiplex protein array and near-infrared (NIR) fluorescence detection. Serum levels of IL-1ra, IL-23, MIF, sCD40 ligand, IP-10 and GRO-α were also determined by ELISA.ResultsAt the early stages of infection, GRO-α and IP-10 expression levels were different in DF compared to DHF patients. Besides, GRO-α was positively correlated with platelet counts and IP-10 was negatively correlated with total protein levels.ConclusionsThese findings suggest that high levels of GRO-α during acute DENV infection may be associated with a good prognosis, while high levels of IP-10 may be a warning sign of infection severity.     

Dengue hemorrhagic fever – A systemic literature review of current perspectives on pathogenesis,prevention and control

BackgroundDengue is an arboviral disease caused by dengue virus. Symptomatic dengue infection causes a wide range of clinical manifestations, from mild dengue fever (DF) to potentially fatal disease, such as dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS). We conducted a literature review to analyze the risks of DHF and current perspectives for DHF prevention and control.MethodsAccording to the PRISMA guidelines, the references were selected from PubMed, Web of Science and Google Scholar database using search strings containing a combination of terms that included dengue hemorrhagic fever, pathogenesis, prevention and control. Quality of references were evaluated by independent reviewers.ResultsDHF was first reported in the Philippines in 1953 and further transmitted to the countries in the region of South-East Asia and Western Pacific. Plasma leakages is the main pathophysiological hallmark that distinguishes DHF from DF. Severe plasma leakage can result in hypovolemic shock. Various factors are thought to impact disease presentation and severity. Virus virulence, preexisting dengue antibodies, immune dysregulation, lipid change and host genetic susceptibility are factors reported to be correlated with the development of DHF. However, the exact reasons and mechanisms that triggers DHF remains controversial. Currently, no specific drugs and licensed vaccines are available to treat dengue disease in any of its clinical presentations.ConclusionThis study concludes that antibody-dependent enhancement, cytokine dysregulation and variation of lipid profiles are correlated with DHF occurrence. Prompt diagnosis, appropriate treatment, active and continuous surveillance of cases and vectors are the essential determinants for dengue prevention and control.     

Transient CD4/CD8 ratio inversion and aberrant immune activation during dengue virus infection

The immune status after dengue virus infection was studied in dengue patients from an outbreak of serotype 3 dengue virus infection in the southern part of Taiwan during November and December 1998. Consecutive blood samples from 29 dengue patients, of whom 21 had dengue fever and 8 had dengue hemorrhagic fever/dengue shock syndrome, were collected, and the immunophenotypes of the peripheral blood mononuclear cells were determined by flow cytometry. The early activation marker CD69 appeared on lymphocytes and monocytes at day 4 after the onset of fever, and declined afterward. However, a transient reverse in the CD4/CD8 ratio occurred at days 6-10 after the onset of fever. The CD4/CD8 ratio inversion was manifested in 10 of 29 dengue patients and was encountered more frequently in dengue hemorrhagic fever/dengue shock syndrome than in dengue fever patients. Analysis of the clinical blood cell count of these 10 cases showed that increase of immature neutrophils developed at fever days 5-6, CD4(dim) or CD8(dim) monocytosis at days 6-7, and atypical lymphocytosis at days 8-10 after the onset of fever. Serum IL-6 was found at either day 7 or day 9-11. The PHA-stimulated T-cell response was depressed as well. These changes in immune parameters indicate aberrant immune activation during dengue virus infection and might be involved in the pathogenesis of dengue virus infection.     

固相酶联免疫测定法检测NS1抗原在登革病毒感染早期诊断中的应用

目的探讨固相酶联免疫测定（ELISA）法检测NS1抗原在登革病毒感染早期诊断中的应用价值。方法选取登革病毒感染早期患者血清171份,非登革病毒感染发热患者血清11份,正常人血清10份,采用ELISA法检测全部192份血清的登革病毒NS1抗原和IgM抗体;采用逆转录-聚合酶链反应-限制性内切酶酶切片段长度多态性分析（RT-PCR-RFLP）技术对发病5 d内的125份血清进行扩增和鉴定分型;并采用C6/36细胞微量培养法对发病第1、2天的41份血清进行登革病毒分离培养。结果登革病毒感染患者发病2 d内、3～5 d以及6～10 d血清NS1抗原的检出率分别是92.7%（38/41）、83.3%（70/84）、10.9%（5/46）;IgM抗体的检出率分别是2.4%（1/41）、51.2%（43/84）、97.8%（45/46）;非登革病毒感染的发热患者及正常人血清中,有1例疟疾患者血清登革病毒IgM抗体呈阳性,NS1抗原无一例阳性。RT-PCR在登革病毒感染患者发病第1、2天和3～5天的检出率分别是85.4%（35/41）、83.3%（70/84）;登革病毒感染患者发病第1、2天血清的病毒分离培养阳性率分别是80.0%（16/20）、38.1%（8/21）,总分离率58.5%（24/41）;RT-PCR-RFLP分型鉴定技术及间接免疫荧光法（IFA）均证实2006年广州流行株为登革Ⅰ型病毒。结论ELISA法检测登革病毒NS1抗原操作技术成熟,且具有敏感性高、特异性好的特点,对登革病毒感染的早期诊断和疫情的早期控制具有重要意义,适合于基层医疗机构常规应用。     

2006年广州地区登革热流行的临床特征

目的总结2006年6～12月间广州地区出现的439例登革热病人的临床特点。方法对2006年我院收治入院的439例登革热病人的临床症状、体征和实验室检查进行回顾性分析。结果2006年的登革热病例主要临床表现为发热（100%）、头痛（75.9%）、全身肌痛（56.7%）、骨痛（42.1%）、恶心呕吐（27.1%）、皮疹（92.9%）;白细胞及血小板减少分别占72.7%和64.7%,血液生化示丙氨酸转氨酶（ALT）升高占57.9%、天门冬氨酸转氨酶（AST）升高占85.2%、低钾血症占44.2%,淋巴结肿大占15.3%,肝肿大占1.4%,未见脾肿大。CD细胞亚群计数CD3下降的占44.5%、CD4下降的占46.2%,CD8下降的占38.7%。所有病例登革热抗体IgM阳性。结论此次广州地区流行的登革热病例,临床表现典型合并有多脏器损害,尤其肝损害较多见,近半数病人出现CD细胞计数明显下降,未出现登革出血热及登革休克综合征。经及时诊断和治疗,预后良好。     

Epidemiology of dengue infections in Kassala, Eastern Sudan

Eighty‐one (71.7%) out of 113 patients had confirmed dengue infection (using ELISA IgM serology) at Kassala, Eastern Sudan during the period of August through November 2010. According to the WHO criteria, dengue fever (DF), dengue hemorrhagic fever (DHF), and dengue shock syndrome (DSS) were observed in 30.9, 58, and 11.1% of these patients, respectively. The mean age of these 81 patients was 25.5 years. Male:female ratio was 1.8:1. Various symptoms including fever (100%), headache (75.3%), vomiting (55.6%), nausea (53.1%), and backache (30.9%) were observed among these patients. Thrombocytopenia (<100/10⁹ platelets/L), and leucopenia (WBC count <4,000 × 10⁹ cells/L) and hemoconcentration (hematocrit >45) were reported in 86.4, 69.1, and 67.9% of the patients, respectively. High alanine aminotransferase (ALT, >65 U/L) and aspartate aminotransferase (AST >37 U/L) were seen in 9.9 and 14.8% of the patients, respectively. There were five (6.1%) deaths, three of them had DHF and the other two patients had DSS. J. Med. Virol. 84:500–503, 2012. © 2011 Wiley Periodicals, Inc.