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PubMed1976

Immunofluorescent staining of the surfaces of lymphocytes in suspension from patients with dengue hemorrhagic fever.

Immunofluorescent staining of lymphocytes suspensions from 55 of 62 patients with dengue hemorrhagic fever was positive for dengue antigen and human beta1C/a-globulin on the surface, from the second ay before shock or subsidence of fever. The percentages of positive staining of both components gradually increased to a maximum on the day of shock or subsidence of fever. B lymphocytes increased during the course of the disease. Neither dengue antigen nor human beta1C/a-globulin was detected on the surface of the lymphocytes from normal controls or patients with other diseases. By double immunofluorescent staining with different colors of fluorochromes, antidengue antibody with fluorescein isothiocyanate and antihuman gamma -globulin or antihuman beta1C/a-globulin with lissamine rhodamine B on the same lymphocytes revealed dengue antigen appearing only on B lymphocytes. The human beta1C/a-globulin and dengue antigen were located on the surface of the same lymphocytes. The pattern of the staining by both components showed fine and coarse irregular granules over the lymphocyte surface. The fluorescent granules seemed to be on the surface but not in the intracellular vacuoles of the lymphocytes.

Alpha-Globulins↗
PubMed1981

Surveillance of dengue hemorrhagic fever cases in Thailand.

A long-term surveillance system is necessary for planning and evaluation of hemorrhagic fever control Dry blood collection and using one dengue 2 antigen have been proved to be an efficient method to detect low and high level of HI antibody, determined to prove dengue infection. The percentage of dengue infection serologically proved from cases reported throughout a 3 year study was 52%. Dengue infected cases were reported outside epidemic period with lowest incidence in January. The majority of dengue proven cases occur at age 6 years. For chikungunya infection, studied in Bangkok metropolis and in this study in 72 provinces shows similar result indicating that chikungunya virus shows no significance in clinical and laboratory study in the surveillance programme. This study provides definite information for the planning and evaluation of hemorrhagic fever control.

Adolescent↗
PubMedDate not supplied

Dengue fever with thrombocytopenia: studies towards defining vulnerability of bleeding.

PURPOSE: To define the period of greater vulnerability of bleeding in patients with Dengue fever in reference to the onset of their constitutional symptoms and the laboratory abnormalities. PATIENTS AND METHODS: In a retrospective study we reviewed the records of all patients admitted to San Pablo Medical Center in 1991 with a diagnosis of Dengue Fever or Hemorrhagic Dengue. All patients with a platelet count of less than 125,000 were included for analysis. The exclusion criteria included the presence of systemic disorders which may influence the platelet count, and patients without documentation regarding the presence of constitutional symptoms suggestive of viral illness. RESULTS: A total of 101 patients were analyzed of which only 74 were included in the study. All patients had fever and chills; skin rash, asthenia and general malaise was seen in over 50% of patients. Over 70% of patients had recovery of their platelet count, and most had their maximal thrombocytopenia within the 5th day and 8th day from the onset of constitutional symptoms. Leukopenia was seen in over 70% of patients with its lowest level within the 5th and 8th day from the onset of the constitutional symptoms. Significantly prolonged partial thromboplastin time was seen in 11% of the patients. Proteinuria was seen in 22% of the patients, 38% of which had it within the first 4 days of the onset of constitutional symptoms and also noted on the 5th and 6th day. Alteration in liver enzymes were noted in 47% of patients, with a maximal severity distributed in all time frames. Hypoalbuminemia was present in 28% of the patients, of these 67% presented within the first 4 days from the onset of constitutional symptoms. The pulse rate was usually normal in spite of the patient's dehydration and fever. CONCLUSIONS: We identified three phases that define the sequence of events seen in the majority of patients with Dengue Fever and Thrombocytopenia. These are: 1. proteinuria and hypoalbuminemia; 2. maximal cytopenia; 3. bradycardia and liver enzyme elevation. We believe this information is useful in the management of patients with Dengue Fever and thrombocytopenia.

Adolescent↗
PubMed1993

[Hemorrhagic dengue with shock in children in French Polynesia].

The dengue fever is an intertropical arbovirus, well known in French Polynesia where the first cases of dengue haemorrhagic fever appeared at the end of 1989. The authors report their experience in the paediatrics ward of the Centre Hospitalier Territorial (CHT) of Papeete, where, from 1990 to 1992, 310 cases of dengue fever have been registered with 79 cases of dengue shock syndrome (25.5%) and 12 deaths (4%). Then, following an epidemiologic summary and a description of how the clinical picture evolves in children, they insist on the fact that the pathophysiology of dengue fever is unknown, and there is no predictive factor; even if, in this study, the young age seems to be factor of severity. Therefore, an epidemiologic survey, and a very close monitoring of each case are necessary, in order to start an aggressive symptomatic treatment as soon as possible. The antivector's fight is still the only prevention against this arbovirosis.

Child↗
PubMed1993

[Dengue. Apropos of 2 cases].

BACKGROUND. Dengue is an acute febrile illness caused by several arthropod-born viruses and characterized by biphasic fever, myalgia or arthralgia, rash, leukopenia and lymphadenopathy. Its diagnosis is based on knowledge of the geographic distribution of dengue viruses. CASE REPORTS Case no 1: A 11 year-old boy suffered from sudden onset of fever accompanied by retro-orbital headache, arthralgia and diffuse myalgia. There was no rash. Hemogram showed: hemoglobin: 11.6 g%; leukocytes: 3,400/mm3 (PMN: 76%); platelets: 190,000/mm3. A diagnosis of viral infection was considered, but, as the boy had recently been to the French West-Indies, a serologic study was performed. This was negative 2 days after the onset of disease and positive (specific IgM for the 4 dengue types), 13 days later. Case no 2: A 7 year-old boy suffered from sudden onset of fever. Severe calf muscle pain 4 days later led to his admission. Creatine phosphokinase activity was very high: 83,100 units (N: 30-120). Hemogram showed: hemoglobin: 11.4 g%; leukocytes: 2,500/mm3 (PMN: 60%); platelets: 124,000/mm3. A diagnosis of acute myositis was considered, but as the patient had recently visited Venezuela, a serologic study was performed. This was negative 8 days after the onset of disease and positive (specific IgM for the 4 dengue types) 16 days later. CONCLUSION. The first case is characteristic of the classical form of dengue fever. The second patient presented with very localized myalgia. The diagnosis in both cases was facilitated by the knowledge that the patient had recently stayed in an endemic area.

Acute Disease↗
PubMed1993

Failure of high-dose methylprednisolone in established dengue shock syndrome: a placebo-controlled, double-blind study.

OBJECTIVE: Steroids are widely used in Thailand and other dengue-endemic countries to treat severe dengue shock syndrome. This study was designed to determine whether a single high dose of methylprednisolone will reduce mortality in children with dengue shock syndrome who did not respond to simple fluid and plasma replacement therapy. METHODS: A prospective, randomized, double-blind, controlled trial was conducted in two hospitals in Khon Kaen Thailand during June to September in 1987 and 1988. Sixty-three children with severe dengue shock syndrome were randomized into two groups; the first group received a single dose of methylprednisolone (30 mg/kg) and the second group received placebo. RESULTS: There was no significant difference in mortality between the two groups (P = .63). The mortality rate was 12.5% (4/32) in the steroid group and 12.9% (4/31) in the group that received placebo. The sequelae at 2 weeks among treatment and control survivors were not significantly different. These two groups were comparable in age, sex, severity of illness, and duration of shock at the outset of the study. The two treatment groups were similar in subsequent hospital course as determined by maximum and minimum hematocrit level and bleeding severity. The numbers of patients in each group who had liver failure and evidence of disseminated intravascular clotting defect were also comparable. Complications such as occurrence of fever after shock, pneumonia, convulsion, cardiac arrest, pulmonary hemorrhage, and positive hemoculture were not significantly different in the treatment and control groups. CONCLUSIONS: A single high dose of methylprednisolone does not reduce mortality in severe dengue shock syndrome which does not respond to conventional critical care.

Adolescent↗
PubMed1995

[The epidemiological situation of dengue in America. The challenges for its surveillance and control].

The recent occurrence of cases of dengue hemorrhagic fever has placed dengue in the agenda of priorities in many countries in the Americas. International efforts directed to the control of the epidemic are supported by the epidemiological surveillance of dengue cases and serotypes while control of the vector is still in need of innovative and more effective measures. These are the challenges to be met by countries in the region, where the laboratory should be incorporated as the key component of the surveillance system. The medical community requires adequate training in the diagnosis and treatment of the severe forms of dengue infection and the community should be deeply involved in the control strategies. Scant resources in the region impose serious constrains to the program which should be comprehensive, intensive and well coordinated. The medical services, the vector control personnel, the surveillance team, the civil authorities and the community have important roles to play in the control of dengue fever.

Aedes↗
PubMed1996

Rapid detection and identification of dengue viruses by polymerase chain reaction (PCR).

A polymerase chain reaction (PCR) method using sets of newly designed primers for rapid detection and simultaneous identification of dengue virus serotypes was developed and tested. The test is based on two sets of primers specific within the envelope (E) and non-structural (NS1) regions of the dengue-virus genome. Two sets of universal primers that bind to two target sequences which are shared by all the four serotypes of the virus within the E and NS1 regions are used. The resulting products are further amplified by another pair of inner or nested universal primers, which also bind to another set of shared sequences within the E and NS1 regions, respectively. The nested PCR of both the E and NS1 regions can detect dengue virus of all the four serotypes at a sensitivity of 1 plaque forming unit (pfu) or less. For the identification of serotypes, a mixture of four pairs of serotype-specific primers, specific to the E region, was used. The primers have been designed to bind to serotype specific sequences within the regions flanked by the outer universal primers, and giving the amplified products of different sizes, each corresponds to one particular serotype (405 bp for Den1, 346 bp for Den2, 196 bp for Den3, and 143 bp for Den4). A protocol has been developed and successfully applied to detect dengue virus in cell-culture supernatants and patients sera. The technique is simple and rapid, capable of not only detecting the dengue virus but also identifying the serotypes of the virus in clinical specimens.

DNA Primers↗
PubMed1997

Dengue fever: a risk to travelers.

An outbreak of dengue fever occurred among a small group of Maryland and Pennsylvania residents following a trip to the British Virgin Islands in January 1996. Dengue fever is a mosquito-borne viral illness that occurs primarily in tropical urban areas. Most dengue infections are benign and self-limited, but some produce severe and fatal hemorrhagic disease. Although dengue is not endemic in the continental United States, travelers may acquire the infection during visits to the tropics. Physicians should consider dengue in the differential diagnosis of a patient with a febrile illness and a history of recent travel to a tropical area. Travelers to endemic areas should be advised to take precautions to prevent mosquito bites.

Adult↗
PubMed2000

Prospective study of the duration and magnitude of viraemia in children hospitalised during the 1996-1997 dengue-2 outbreak in French Polynesia.

The magnitude and duration of viraemia in children admitted to the hospital with dengue was studied during a dengue 2 outbreak in French Polynesia in 1996-1997. Forty-nine patients from whom at least 3 plasma samples were available were included in the study. Based on analysis of IgG-ELISA and haemagglutination inhibition assay, 21 of these were primary and 28 were secondary infections. According to World Health Organization criteria, 42 were dengue fever and 7 were dengue haemorrhagic fever. Virus was detectable by reverse transcription-PCR in all patients for at least the first 3 days of the onset of fever, but was never detected after the 6th day (mean duration = 4.4 days). Plasma virus titers ranged from 1.7-5.6 Log(10) TCID(50)/ml. A significant difference was not observed in the magnitude and duration of viraemia in patients with primary versus secondary infections. The severity of the illness, however, was correlated with both criteria.

Antibodies, Viral↗
PubMed2004

Emergence and continued circulation of dengue-2 (genotype IV) virus strains in northern India.

Dengue (DEN) is an acute mosquito borne viral disease of mankind. Off late it has become an important public health concern in Southeast Asia. Although, all the four known dengue virus serotypes (DEN-1 to 4) are reported from time to time, in the recent past, DEN-2 has emerged as the predominant type, being the causative agent of several outbreaks of dengue fever (DF) and dengue haemorrhagic fever (DHF) in India. To elucidate the true molecular epidemiology of these viruses, we have sequenced C-prM gene junction (454 nucleotides) of 11 DEN-2 viruses directly from patient serum. The C-prM gene junction was amplified initially by reverse transcription-polymerase chain reaction followed by automated DNA sequencing. These sequences provide unique information with regard to molecular epidemiology when compared to other DEN-2 sequences from diverse geographic origins. The sequence analysis revealed that most of the mutations in this region remained silent, except a few at the carboxy-terminal of the capsid. Reported phylogenetic analysis classifies DEN-2 viruses into five distinct genotypes. The Gwalior DEN-2 viruses, included in the present study were classified into genotype-IV, and were found to be most closely related to Delhi 1996 DEN-2 viruses and FJ 10/11 strains prevalent in the Fujian state of China. However, two earlier Indian isolates of DEN-2 were classified into genotype-V. The present study indicates that genotype V of DEN-2 has been replaced by genotype IV during the past decade, which continues to circulate silently in north India, and have the potential to reemerge and cause major epidemics of DF and DHF.

Base Sequence↗
PubMed2004

Nucleotide sequence and phylogenetic analyses of dengue type 2 virus isolated in the Dominican Republic.

Dengue virus infection has been recognized as an important public health problem in the Dominican Republic in the last decade. Complete genomic sequences of three strains of dengue type 2 (DEN-2) virus, DR23/01 and DR31/01 isolated from dengue fever (DF) patients, and DR59/01 isolated from a dengue hemorrhagic fever (DHF) patient, all with primary infection, in the Dominican Republic in 2001, have been established. This achievement constitutes the first genomic characterization of DEN-2 strains from the Dominican Republic. No amino acid differences were observed between the strains isolated from DHF and DF patients. They exhibited extensive homology with the strain from La Martinique, French West Indies. Although phylogenetic analysis was suggestive of their Southeast Asiatic origin, Dominican Republic strains and other Caribbean strains from La Martinique and Jamaica showed 26 amino acid changes that differed from both the Southeast Asia and native American genotypes.

Base Sequence↗
PubMed1964

DENGUE-TYPE VIRUSES ISOLATED IN SINGAPORE.

A dengue-like illness with marked haemorrhagic manifestations appeared in Singapore in 1960. Its similarity in many respects to the haemorrhagic fevers of Thailand and the Philippines led to its being described as "Singapore haemorrhagic fever".This paper describes the isolation and identification of dengue-type viruses from patients in Singapore between 1960 and 1962. In addition to the conventional complement-fixation and neutralization tests, a new test, called the "sensitized erythrocyte agglutination test", was employed; this test method is described.Altogether 21 dengue-type viruses were isolated, including dengue types 1, 2 and 4. Chikungunya virus, prominent in the Thailand disease, was not detected.The author suggests that study of the epidemiology of haemorrhagic fevers in South-East Asia would cast further light on the transmission of arboviruses.

Arboviruses↗
PubMed2005

[Inoculation of plasmid expressing the dengue 2 NS1 gene elicits immunity in mice].

OBJECTIVE: To study cellular and humoral immune responses to NS1 protein in mice inoculated intramuscularly with recombinant plasmid expressing dengue 2 NS1 gene. METHODS: The eukaryotic expressing plasmid pCNX2-NS1 was injected into tibialis anterior muscle in mice. The mice were subsequently boosted with the same dose and same method twice after the initial inoculation. The mice were killed at four-week intervals and their serum and spleen cells were harvested for further test. RESULTS: Dengue 2 antibodies were detectable in the sera from inoculated animals four weeks after the last boost. The changes of CD4+ T lymphocyte and CD8+ T lymphocyte were also determined by flow cytometry. CONCLUSION: The recombinant plasmid containing dengue 2 NS1 genes is immunogenic in intramuscularly inoculated mice. The vaccinated mice produced dengue-2 specific and long lasting immunity.

Animals↗
PubMedDate not supplied

[Epidemic outbreak of dengue virus 2/Jamaica genotype in Bolivia].

OBJECTIVE: To confirm an epidemic outbreak of Dengue virus in the city of Santa Cruz, Bolivia, and to determine the serotype of the virus, to estimate the rate of attack and the proportion of symptomatic infections. MATERIAL AND METHODS: In March 1997, a seroepidemiological survey was conducted with random sampling in a central district of the city of Santa Cruz, Bolivia. Information on recent acute illness and febrile episodes was gathered, and venous blood samples were obtained. Levels of antidengue IgM were determined by MAC Elisa and the virus was typified with RT-PCR. RESULTS: IgM antibodies were detected in 6.5% of adults (CI 95% 3.4-9.6) and 5.1% of children (CI 95% 2.0-8.2). Circulating virus was identified as Dengue serotype 2, subgroup Jamaica. Less than half of the infected children experienced a symptomatic infection compared to almost 90% of adults. CONCLUSIONS: The estimated attack rates are compatible with a Dengue epidemic outbreak in Santa Cruz. The introduction of the serotype 2/ subgroup Jamaica virus into the country increases the risk of hemorrhagic Dengue.

Adolescent↗
PubMed1994

A shortened dengue IgM capture ELISA using simultaneous incubation of antigen and peroxidase-labeled monoclonal antibody.

A shortened IgM capture ELISA for the detection of dengue IgM antibodies using simultaneous incubation of antigen and peroxidase-labeled monoclonal antibody was described. The shortened two-step assay was compared with the four-step IgM capture ELISA on sera from dengue patients confirmed by the hemagglutination inhibition (HI) test. When paired acute and convalescent sera were tested, the shortened ELISA showed 100% agreement with HI results. It detected dengue IgM antibodies in the acute sera of 66% of patients with a primary dengue infection, 60% of patients with a secondary infection, and 98% of patients with a presumptive secondary infection. When the results of 151 dengue patients were combined, 75% of the acute sera were positive by the shortened IgM capture ELISA.

Journal Article↗
PubMed2006

Substrate specificity of recombinant dengue 2 virus NS2B-NS3 protease: influence of natural and unnatural basic amino acids on hydrolysis of synthetic fluorescent substrates.

A recombinant dengue 2 virus NS2B-NS3 protease (NS means non-structural virus protein) was compared with human furin for the capacity to process short peptide substrates corresponding to seven native substrate cleavage sites in the dengue viral polyprotein. Using fluorescence resonance energy transfer peptides to measure kinetics, the processing of these substrates was found to be selective for the Dengue protease. Substrates containing two or three basic amino acids (Arg or Lys) in tandem were found to be the best, with Abz-AKRRSQ-EDDnp being the most efficiently cleaved. The hydrolysis of dipeptide substrates Bz-X-Arg-MCA where X is a non-natural basic amino acid were also kinetically examined, the best substrates containing aliphatic basic amino acids. Our results indicated that proteolytic processing by dengue NS3 protease, tethered to its activating NS2B co-factor, was strongly inhibited by Ca2+ and kosmotropic salts of the Hofmeister's series, and significantly influenced by substrate modifications between S4 and S6'. Incorporation of basic non-natural amino acids in short peptide substrates had significant but differential effects on Km and k(cat), suggesting that further dissection of their influences on substrate affinity might enable the development of effective dengue protease inhibitors.

Amino Acids, Basic↗
PubMed1996

Imported Dengue in The Netherlands.

Background: The clinical manifestations of imported dengue in adult Western travelers were analyzed. Methods: The charts of all adult patients with dengue, who were seen in an academic hospital in The Netherlands from 1985 to 1994, were studied. Results: Dengue was confirmed in 34 patients by a fourfold rise in complement fixation antibody titer or presence of IgM. Twelve probable cases with a single high titer in the complement fixation test were found. Annual numbers have increased since 1989. Most cases were imported from Asia. Malaise (98%), fever (96%), headache (64%), chills (58%), muscle pain (53%) and arthralgia (44%) were the main symptoms. A noncharacteristic skin eruption was seen in 84%. Leukocytopenia and thrombocytopenia were found in 87% and 75%, respectively during the first week, and elevated serum aminotransferase activities were noted in 75%. Only two patients developed dengue hemorrhagic fever. Conclusions: Dengue in Western travelers is usually a relatively mild disease. Leukocytopenia, thrombocytopenia, and elevation of serum aminotransferase activities are common laboratory features.

Journal Article↗