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Fulltext Development of standard clinical endpoints for use in dengue interventional trials

Tomashek KM, Wills B, See Lum LC, Thomas L, Durbin A, Leo YS, et al.

PLoS Negl Trop Dis, 2018 10;12(10):e0006497.
PMID: 30286085 DOI: 10.1371/journal.pntd.0006497

Dengue is a major public health problem worldwide. Although several drug candidates have been evaluated in randomized controlled trials, none has been effective and at present, early recognition of severe dengue and timely supportive care are used to reduce mortality. While the first dengue vaccine was recently licensed, and several other candidates are in late stage clinical trials, future decisions regarding widespread deployment of vaccines and/or therapeutics will require evidence of product safety, efficacy and effectiveness. Standard, quantifiable clinical endpoints are needed to ensure reproducibility and comparability of research findings. To address this need, we established a working group of dengue researchers and public health specialists to develop standardized endpoints and work towards consensus opinion on those endpoints. After discussion at two working group meetings and presentations at international conferences, a Delphi methodology-based query was used to finalize and operationalize the clinical endpoints. Participants were asked to select the best endpoints from proposed definitions or offer revised/new definitions, and to indicate whether contributing items should be designated as optional or required. After the third round of inquiry, 70% or greater agreement was reached on moderate and severe plasma leakage, moderate and severe bleeding, acute hepatitis and acute liver failure, and moderate and severe neurologic disease. There was less agreement regarding moderate and severe thrombocytopenia and moderate and severe myocarditis. Notably, 68% of participants agreed that a 50,000 to 20,000 mm3 platelet range be used to define moderate thrombocytopenia; however, they remained divided on whether a rapid decreasing trend or one platelet count should be case defining. While at least 70% agreement was reached on most endpoints, the process identified areas for further evaluation and standardization within the context of ongoing clinical studies. These endpoints can be used to harmonize data collection and improve comparability between dengue clinical trials.

Matched MeSH terms: Dengue/diagnosis
Development of a dot enzyme immunoassay for dengue 3: a sensitive method for the detection of antidengue antibodies

Cardosa MJ, Hooi TP, Shaari NS

J Virol Methods, 1988 Oct;22(1):81-8.
PMID: 3058737

Partially purified DEN3 virus was used as antigen in a sensitive dot enzyme immunoassay (DEIA) for the detection of antibodies to flavivirus antigens. We describe here the method used to prepare and optimise the antigen-bearing nitrocellulose membranes and present the results obtained from screening 20 acute phase sera from patients shown to have had recent dengue infections by the haemagglutination inhibition (HI) test. Sixteen pairs of acute and convalescent sera from dengue-negative patients had no detectable antibody to dengue virus by HI. These were shown to have no antibody detectable by DEIA. Sera positive for dengue antibodies by HI had DEIA titers ranging from 10 to several thousand times greater than the titers detected by HI.

Matched MeSH terms: Dengue/diagnosis
Detection of specific IgM in dengue infection

Lam SK, Devi S, Pang T

Southeast Asian J. Trop. Med. Public Health, 1987 Dec;18(4):532-8.
PMID: 3329413

A modification of the IgM-capture ELISA which can provide an early diagnosis for dengue infection is presented. The test is technically simple compared to HI and appears to be more sensitive. It has the advantage over HIT for the detection of specific IgM in that it is more sensitive and the reading of the result is not subjective. There is the possibility of the test being able to replace HI and HIT in the future.

Matched MeSH terms: Dengue/diagnosis*
Fulltext Detection of dengue viruses using reverse transcription-loop-mediated isothermal amplification

Teoh BT, Sam SS, Tan KK, Johari J, Danlami MB, Hooi PS, et al.

BMC Infect Dis, 2013;13:387.
PMID: 23964963 DOI: 10.1186/1471-2334-13-387

BACKGROUND: Early and rapid detection of dengue virus (DENV) infection during the febrile period is crucial for proper patient management and prevention of disease spread. An easy to perform and highly sensitive method is needed for routine implementation especially in the resource-limited rural healthcare settings where dengue is endemic.
METHODS: A single-tube reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay with a set of nine primers was developed for the detection of all four DENV serotypes and their different genotypes. The sensitivity and specificity of the RT-LAMP were evaluated. The clinical applicability of RT-LAMP assay for detection of DENV RNA was assessed in a total of 305 sera of clinically-suspected dengue patients. The test results of RT-LAMP were statistically compared to those of quantitative reverse transcription-polymerase chain reaction (qRT-PCR), IgM- and IgG-capture enzyme-linked immunosorbent assays (ELISA).
RESULTS: Acute DENV infection was confirmed in 171 samples (n = 305); 43.3% (74/171) and 46.8% (80/171) of the samples were positive for DENV using RT-LAMP and qRT-PCR, respectively. The combination of RT-LAMP with the dengue IgM and IgG ELISA increased detection of acute DENV infection to 97.7% (167/171), in comparison to only 70.8% (121/171) when dengue IgM and IgG ELISA alone were used. The RT-LAMP assays showed high concordance (κ = 0.939) with the qRT-PCR. The RT-LAMP assay detected up to 10 copies of virus RNA within an hour but 100% reproducibility (12/12) was achieved with 100 copies. There was no cross reactivity of RT-LAMP with other closely related arboviruses.
CONCLUSION: The RT-LAMP assay developed in this study is sensitive, specific and simple to perform. The assay improved the detection of dengue when used in combination with serological methods.

Matched MeSH terms: Dengue/diagnosis*
Fulltext Detection of dengue using PAMAM dendrimer integrated tapered optical fiber sensor

Mustapha Kamil Y, Al-Rekabi SH, Yaacob MH, Syahir A, Chee HY, Mahdi MA, et al.

Sci Rep, 2019 09 17;9(1):13483.
PMID: 31530893 DOI: 10.1038/s41598-019-49891-7

The exponential escalation of dengue cases has indeed become a global health crisis. This work elaborates on the development of a biofunctionalized tapered optical fiber (TOF) based sensor with the integration of polyamidoamine (PAMAM) dendrimer for the detection of dengue E protein. The dimension of the TOF generated an evanescent field that was sensitive to any changes in the external medium while the integration of PAMAM promoted more adhesion of bio-recognition molecules; anti-DENV II E protein antibodies; that were complementary to the targeted protein. This in return created more active sites for the absorption of DENV II E proteins onto the tapered region. The resolution and detection limit of the sensor are 19.53 nm/nM and 1 pM, respectively with Kd = 1.02 × 10-10 M.

Matched MeSH terms: Dengue/diagnosis*
Fulltext Detection of Wolbachia from field collected Aedes albopictus Skuse in Malaysia

Noor Afizah A, Roziah A, Nazni WA, Lee HL

Indian J Med Res, 2015 Aug;142(2):205-10.
PMID: 26354218 DOI: 10.4103/0971-5916.164259

Wolbachia-based vector control strategies have been proposed as a mean to augment the existing measures for controlling dengue vector. Prior to utilizing Wolbachia in novel vector control strategies, it is crucial to understand the Wolbachia-mosquito interactions. Many studies have only focused on the prevalence of Wolbachia in female Aedes albopictus with lack of attention on Wolbachia infection on the male Ae. albopictus which also affects the effective expression of Wolbachia induced- cytoplasmic incompatibility (CI). In this study, field surveys were conducted to screen for the infection status of Wolbachia in female and male Ae. albopictus from various habitats including housing areas, islands and seashore.

Matched MeSH terms: Dengue/diagnosis
Fulltext Dengue related maculopathy and foveolitis

Juanarita J, Azmi MN, Azhany Y, Liza-Sharmini AT

Asian Pac J Trop Biomed, 2012 Sep;2(9):755-6.
PMID: 23570008 DOI: 10.1016/S2221-1691(12)60223-8

A 24 year-old Malay lady presented with high grade fever, myalgia, generalized rashes, severe headache and was positive for dengue serology test. Her lowest platelet count was 45 × 10(9) cells/L. She complained of sudden onset of painlessness, profound loss of vision bilaterally 7 days after the onset of fever. On examination, her right eye best corrected vision was 6/30 and left eye was 6/120. Her anterior segment examination was unremarkable. Funduscopy revealed there were multiple retinal haemorrhages found at posterior pole of both fundi and elevation at fovea area with subretinal fluid. Systemic examination revealed normal findings except for residual petechial rashes. She was managed conservatively. Her vision improved tremendously after 2 months. The retinal hemorrhages and foveal elevation showed sign of resolving. Ocular manifestations following dengue fever is rare. However, bilateral visual loss can occur if both fovea are involved.

Matched MeSH terms: Dengue/diagnosis
Fulltext Dengue haemorrhagic fever with unusual prolonged thrombocytopaenia

Kamil SM, Mohamad NH, Narazah MY, Khan FA

Singapore Med J, 2006 Apr;47(4):332-4.
PMID: 16572249

We describe a case of dengue haemorrhagic fever with prolonged thrombocytopaenia. A 22-year-old Malay man with no prior illness presented with a history of fever and generalised macular rash of four days duration. Initial work-up suggested the diagnosis of dengue haemorrhagic fever based on thrombocytopaenia and positive dengue serology. Patient recovered from acute illness by day ten, and was discharged from the hospital with improving platelet count. He was then noted to have declining platelet count on follow-up and required another hospital admission on day 19 of his illness because of declining platelet count. The patient remained hospitalised till day 44 of his illness and managed with repeated platelet transfusion and supportive care till he recovered spontaneously.

Matched MeSH terms: Severe Dengue/diagnosis*
Dengue haemorrhagic fever in Malaysia: a review

George R

Southeast Asian J. Trop. Med. Public Health, 1987 Sep;18(3):278-83.
PMID: 3324361

The historical background, epidemiology and changing pattern of clinical disease as seen in Malaysia is reviewed. The preliminary results of the longitudinal study of epidemiology of dengue in Malaysia is also presented. Studies led by Rudnick et al. over some 18 years have established that the disease is endemically transmitted by both Aedes aegypti and Aedes albopictus causing illnesses ranging from mild febrile episodes through classical dengue fever, dengue haemorrhagic fever and the dengue shock syndrome. The first epidemic occurred in 1962 in Penang, and the second major epidemic in 1974 in Selangor. From then on epidemics seem to occur every 4 years, i.e. 1978, and then in 1982. With increasing number of cases being seen from the end of 1985 and in 1986, and with the increasing numbers of positive virus isolates, another epidemic may occur this year. Though in the early years, mainly children were affected, recently more cases are being seen in 16-30 years age group. There is also a changing pattern in the clinical presentation of the cases. The clinician has to be aware of the various modes of presentation of this sinister disease. A high index of suspicion is needed for early diagnosis, as management is mainly symptomatic and there is no specific drug as yet to combat the shock and bleeding manifestations.

Matched MeSH terms: Dengue/diagnosis
Dengue encephalitis: an entity now common in dengue-prone regions

Matlani M, Chakravarti A, Rawal A, Kashyap B, Gurtoo A

Trop Doct, 2009 Apr;39(2):115-6.
PMID: 19299303 DOI: 10.1258/td.2008.080257

As well as dengue fever (DF) and dengue haemorrhagic fever-dengue shock syndrome (DHF/DSS), other atypical manifestations of dengue virus infection have also been reported. The frequency of CNS involvement in dengue remains unknown, although isolated cases with neurological manifestations have been reported in Southeast Asia, Malaysia, Burma, Puerto Rico and India. We present two cases of encephalitis associated with DF and DHF from New Delhi, India.

Matched MeSH terms: Dengue/diagnosis*
Dengue and dengue haemorrhagic fever, Malaysia

Wkly. Epidemiol. Rec., 1998 Jun 12;73(24):182-3.
PMID: 9652206
Matched MeSH terms: Dengue/diagnosis; Severe Dengue/diagnosis
Dengue Maculopathy Associated with Choroidopathy and Pseudohypopyon: A Case Series

Ng CWK, Tai PY, Oli Mohamed S

Ocul Immunol Inflamm, 2018;26(5):666-670.
PMID: 27929712 DOI: 10.1080/09273948.2016.1254804
Matched MeSH terms: Dengue/diagnosis
Concomitant dengue fever in Varicella zoster infection - A rare presentation

Shaiful Ehsan SM, Iskandar FO, Mohd Ashraf AR

Med J Malaysia, 2019 08;74(4):347-348.
PMID: 31424049

Varicella zoster infection is one of the self-limiting viral infections during childhood and dengue fever is an endemic infection in Malaysia, which commonly occurs in the form of nonspecific febrile illness at the initial stage. It is rare for the two viral infections to occur simultaneously. A case of dengue fever without warning sign in a five-year old girl was reported, with early symptoms of fever and vesicular rashes. She was clinically diagnosed with varicella zoster infection during the first visit. Surprisingly, she remained febrile even on day six of illness despite no new vesicular lesions on her skin. Due to suspicion of another infection, follow-up investigation was done and revealed isolated thrombocytopenia. This finding was confirmed with positive NS1Ag. A case of rare dengue fever concomitant with varicella zoster infection was reported.

Matched MeSH terms: Dengue/diagnosis*
Fulltext Comparison of an IgM capture ELISA with a dot enzyme immunoassay for laboratory diagnosis of dengue virus infections

Cardosa MJ, Zuraini I

Southeast Asian J. Trop. Med. Public Health, 1991 Sep;22(3):337-40.
PMID: 1818383

This study describes the use of an IgM capture ELISA using cell culture derived antigens and a polyclonal rabbit antiflavivirus antisera for the detection of dengue positive cases. The IgM capture ELISA is compared with the dot enzyme immunoassay and the results are discussed in the context of dengue endemicity.

Matched MeSH terms: Dengue/diagnosis
Fulltext Comparing the usefulness of the 1997 and 2009 WHO dengue case classification: a systematic literature review

Horstick O, Jaenisch T, Martinez E, Kroeger A, See LL, Farrar J, et al.

Am J Trop Med Hyg, 2014 Sep;91(3):621-34.
PMID: 24957540 DOI: 10.4269/ajtmh.13-0676

The 1997 and 2009 WHO dengue case classifications were compared in a systematic review with 12 eligible studies (4 prospective). Ten expert opinion articles were used for discussion. For the 2009 WHO classification studies show: when determining severe dengue sensitivity ranges between 59-98% (88%/98%: prospective studies), specificity between 41-99% (99%: prospective study) - comparing the 1997 WHO classification: sensitivity 24.8-89.9% (24.8%/74%: prospective studies), specificity: 25%/100% (100%: prospective study). The application of the 2009 WHO classification is easy, however for (non-severe) dengue there may be a risk of monitoring increased case numbers. Warning signs validation studies are needed. For epidemiological/pathogenesis research use of the 2009 WHO classification, opinion papers show that ease of application, increased sensitivity (severe dengue) and international comparability are advantageous; 3 severe dengue criteria (severe plasma leakage, severe bleeding, severe organ manifestation) are useful research endpoints. The 2009 WHO classification has clear advantages for clinical use, use in epidemiology is promising and research use may at least not be a disadvantage.

Matched MeSH terms: Dengue/diagnosis; Severe Dengue/diagnosis
Fulltext Colorimetric Detection of Dengue by Single Tube Reverse-Transcription-Loop-Mediated Isothermal Amplification

Lau YL, Lai MY, Teoh BT, Abd-Jamil J, Johari J, Sam SS, et al.

PLoS One, 2015;10(9):e0138694.
PMID: 26384248 DOI: 10.1371/journal.pone.0138694

Dengue is usually diagnosed by isolation of the virus, serology or molecular diagnostic methods. Several commercial kits for the diagnosis of dengue are existing, but concerns have arisen regarding to the affordability and performance characteristics of these kits. Hence, the loop-mediated isothermal amplification (LAMP) is potentially ideal to be used especially in resource limited environments. Serum was collected from healthy donors and patients diagnosed with dengue infection. RNA extracted from the serum samples were tested by reverse-transcription-LAMP assay developed based on 3'-NCR gene sequences for DENV 1-4. Results were interpreted by a turbidity meter in real time or visually at the end of the assay. Sensitivity and specificity of RT-LAMP results were calculated and compared to qRT-PCR and ELISA. RT-LAMP is highly sensitive with the detection limit of 10 RNA copies for all serotypes. Dengue virus RNA was detected in all positive samples using RT-LAMP and none of the negative samples within 30-45 minutes. With continuing efforts in the optimization of this assay, RT-LAMP may provide a simple and reliable test for detecting DENV in areas where dengue is prevalent.

Matched MeSH terms: Dengue/diagnosis*
Fulltext Cohorting Dengue Patients Improves the Quality of Care and Clinical Outcome

Lum LC, Syed Omar SF, Sri La Sri Ponnampalavanar S, Tan LH, Sekaran SD, Kamarulzaman A

PLoS Negl Trop Dis, 2015 Jun;9(6):e0003836.
PMID: 26047325 DOI: 10.1371/journal.pntd.0003836

INTRODUCTION: The increasing incidence of dengue among adults in Malaysia and other countries has important implications for health services. Before 2004, in order to cope with the surge in adult dengue admissions, each of the six medical wards in a university hospital took turns daily to admit and manage patients with dengue. Despite regular in-house training, the implementation of the WHO 1997 dengue case management guidelines by the multiple medical teams was piecemeal and resulted in high variability of care. A restructuring of adult dengue inpatient service in 2004 resulted in all patients being admitted to one ward under the care of the infectious disease unit. Hospital and Intensive Care Unit admission criteria, discharge criteria and clinical laboratory testing were maintained unchanged throughout the study period.
OBJECTIVES: To evaluate the impact of cohorting adult dengue patients on the quality of care and the clinical outcome in a university hospital in Malaysia.
METHODS: A pre (2003) and post-intervention (2005-6) retrospective study was undertaken.
INTERVENTION: Cohorting all dengue patients under the care of the Infectious Disease team in a designated ward in 2004.
RESULTS: The number of patients enrolled was 352 in 2003, 785 in 2005 and 1158 in 2006. The evaluation and detection of haemorrhage remained high (>90%) and unchanged throughout the study period. The evaluation of plasma leakage increased from 35.4% pre-intervention to 78.8% post-intervention (p = <0.001) while its detection increased from 11.4% to 41.6% (p = <0.001). Examination for peripheral perfusion was undertaken in only 13.1% of patients pre-intervention, with a significant increase post-intervention, 18.6% and 34.2% respectively, p = <0.001. Pre-intervention, more patients had hypotension (21.5%) than detected peripheral hypoperfusion (11.4%), indicating that clinicians recognised shock only when patients developed hypotension. In contrast, post-intervention, clinicians recognised peripheral hypoperfusion as an early sign of shock. The highest haematocrit was significantly higher post-intervention but the lowest total white cell counts and platelet counts remained unchanged. A significant and progressive reduction in the use of platelet transfusions occurred, from 21.7% pre-intervention to 14.6% in 2005 and 5.2% in 2006 post-intervention, p<0.001. Likewise, the use of plasma transfusion decreased significantly from 6.1% pre-intervention to 4.0% and 1.6% in the post-intervention years of 2005 and 2006 respectively, p<0.001. The duration of intravenous fluid therapy decreased from 3 days pre-intervention to 2.5 days (p<0.001) post-intervention; the length of hospital stay reduced from 4 days pre- to 3 days (p<0.001) post-intervention and the rate of intensive care admission from 5.8% pre to 2.6% and 2.5% post-intervention, p = 0.005.
CONCLUSION: Cohorting adult dengue patients under a dedicated and trained team of doctors and nurses led to a substantial improvement in quality of care and clinical outcome.

Matched MeSH terms: Dengue/diagnosis
Fulltext Co-infection of dengue virus and chikungunya virus in two patients with acute febrile illness

Nayar SK, Noridah O, Paranthaman V, Ranjit K, Norizah I, Chem YK, et al.

Med J Malaysia, 2007 Oct;62(4):335-6.
PMID: 18551940 MyJurnal

During an outbreak of chikungunya in a dengue hyperendemic area within the Kinta district of Perak, two patients with acute febrile illness were laboratory confirmed to have co-infection of both dengue and chikungunya viruses in their blood. The concomitant presence of two types of viruses transmitted by the same vector in a susceptible population contributed to the resultant event. A good understanding of virus vector ecology in association with population dynamics and wider application of improved laboratory techniques by using different cell-lines suited for optimal replication of each type of virus and the correct utilization of powerful molecular techniques will enhance accurate diagnosis of these infectious diseases.

Matched MeSH terms: Dengue/diagnosis*
Fulltext Clinico-laboratory spectrum of dengue viral infection and risk factors associated with dengue hemorrhagic fever: a retrospective study

Mallhi TH, Khan AH, Adnan AS, Sarriff A, Khan YH, Jummaat F

BMC Infect Dis, 2015 Sep 30;15:399.
PMID: 26423145 DOI: 10.1186/s12879-015-1141-3

BACKGROUND: The incidence of dengue is rising steadily in Malaysia since the first major outbreak in 1973. Despite aggressive measures taken by the relevant authorities, Malaysia is still facing worsening dengue crisis over the past few years. There is an urgent need to evaluate dengue cases for better understanding of clinic-laboratory spectrum in order to combat this disease.
METHODS: A retrospective analysis of dengue patients admitted to a tertiary care teaching hospital during the period of six years (2008 - 2013) was performed. Patient's demographics, clinical and laboratory findings were recorded via structured data collection form. Patients were categorized into dengue fever (DF) and dengue hemorrhagic fever (DHF). Appropriate statistical methods were used to compare these two groups in order to determine difference in clinico-laboratory characteristics and to identify independent risk factors of DHF.
RESULTS: A total 667 dengue patients (30.69 ± 16.13 years; Male: 56.7 %) were reviewed. Typical manifestations of dengue like fever, myalgia, arthralgia, headache, vomiting, abdominal pain and skin rash were observed in more than 40 % patients. DHF was observed in 79 (11.8 %) cases. Skin rash, dehydration, shortness of breath, pleural effusion and thick gall bladder were more significantly (P 40 years (OR: 4.1, P

Matched MeSH terms: Dengue/diagnosis*; Severe Dengue/diagnosis*
Clinicians' diagnostic practice of dengue infections

Ng CF, Lum LC, Ismail NA, Tan LH, Tan CP

J Clin Virol, 2007 Nov;40(3):202-6.
PMID: 17928264 DOI: 10.1016/j.jcv.2007.08.017

BACKGROUND: Difficulties in the classification of dengue infection have been documented. Such difficulties could be due to the low awareness of the World Health Organization diagnostic guidelines among clinicians.
OBJECTIVE: To study the diagnostic practices of clinicians in classifying patients as dengue fever (DF) or dengue haemorrhagic fever (DHF)/dengue shock syndrome (DSS) at the time of discharge during an outbreak.
METHODS: A prospective descriptive study of clinical features and disease classification in adult and pediatric dengue patients in the University of Malaya Medical Centre.
RESULTS: Five hundred and twenty adult and 191 pediatric patients were enrolled. Thrombocytopenia and evidence of plasma leakage were present in 8% of adult and 19% of pediatric patients. Of these, 93% and 49%, respectively, were given the discharge diagnoses of DF instead of DHF/DSS. Hemoconcentration, serous effusion and thrombocytopenia were not recognized in clinicians' discharge diagnosis of DHF/DSS for adult patients. The receiver operating characteristic (ROC) curve suggested a lack of consistency in the use of WHO guidelines in establishing DHF/DSS in adult patients, while implying otherwise for pediatric patients.
CONCLUSION: DHF/DSS is an under-recognized condition by clinicians managing these patients. This can affect the case fatality rate of DHF/DSS and the economic burden of the disease. The lack of awareness in disease manifestations especially plasma leakage, can lead to delayed recognition of DHF/DSS.
Study site: Outpatient department and inpatients, adult medical and pediatric wards, University Malaya Medical Center (UMMC), Kuala Lumpur, Malaysia

Matched MeSH terms: Dengue/diagnosis*; Severe Dengue/diagnosis*

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