Performance of commercial dengue NS1 ELISA and molecular analysis of NS1 gene of dengue viruses obtained during surveillance in Indonesia

Dengue disease is still a major health problem in Indonesia. Surabaya, the second largest city in the country, is endemic for dengue. We report here on dengue disease in Surabaya, investigating the clinical manifestations, the distribution of dengue virus (DENV) serotypes, and the relationships between clinical manifestations and the genetic characteristics of DENV. A total of 148 patients suspected of having dengue were recruited during February-August 2012. One hundred one (68%) of them were children, and 47 (32%) were adults. Dengue fever (DF) and Dengue hemorrhagic fever (DHF) were equally manifested in all of the patients. We performed DENV serotyping on all of the samples using real-time RT-PCR. Of 148, 79 (53%) samples were detected as DENV positive, with DENV-1 as the predominant serotype (73%), followed by DENV-2 (8%), DENV-4 (8%), and DENV-3 (6%), while 5% were mixed infections. Based on the Envelope gene sequences, we performed phylogenetic analyses of 24 isolates to genotype the DENV circulating in Surabaya in 2012, and the analysis revealed that DENV-1 consisted of Genotypes I and IV, DENV-2 was of the Cosmopolitan genotype, the DENV-3 viruses were of Genotype I, and DENV-4 was detected as Genotype II. We correlated the infecting DENV serotypes with clinical manifestations and laboratory parameters; however, no significant correlations were found. Amino acid analysis of Envelope protein did not find any unique mutations related to disease severity.

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“…These low numbers were in accordance with previous studies describing the low sensitivity of NS1 detection in Indonesia [49,50]. …”

Section: Discussionsupporting

confidence: 93%

Clinical and virological characteristics of dengue in Surabaya, Indonesia

et al. 2017

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“…Interestingly, NS1 antigen was detected in serum samples of DENV-4 patients, but not in urine samples. Using the present ELISA method, other investigators have demonstrated that the NS1 antigen-positive rate of serum samples from DENV-4 patients (19z) was lower than that of other DENV serotypes (DENV1, 67z; DENV-2, 68z; DENV-3, 69z) (30). Further studies using a higher number of urine samples obtained from DENV-4 patients are expected to address the discrepancies in NS1 antigen detection rates between serotypes.…”

Section: Discussionmentioning

confidence: 99%

Detecting Dengue Virus Nonstructural Protein 1 (NS1) in Urine Samples Using ELISA for the Diagnosis of Dengue Virus Infection

et al. 2015

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SUMMARY:Dengue virus (DENV) infection is a serious global health threat. For the surveillance and control of dengue, there is a need for robust diagnostic tools that are relatively easy to use and reliable in various clinical settings. We investigated the applicability of NS1 antigen detection in urine samples for the diagnosis of DENV. About 118 urine samples, obtained from 96 dengue patients at various phases of disease, were used for this study. NS1 antigen was detected by ELISA in the urine samples obtained from patients after 2-17 days of disease onset. Positive detection rates of NS1 antigen ranged between 13-43z. Based on real-time RT-PCR, positive detection rates of viral genome in the urine samples ranged between 20-33z on days 0 to 15. On days 11 to 15 after the disease onset, NS1 antigen was detected at similar rates in serum and urine samples. Additionally, NS1 antigen was detected in 2 urine samples, but not in the serum samples, on days 7 and 16 after the onset of the disease. The results confirm the applicability of NS1 antigen detection in urine samples using ELISA to diagnose acute DENV infection and suggests that the assay is potentially useful when only limited amounts of serum samples are available and in limited resource settings.

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“…Results from these studies showed that these tests had lower sensitivity for the detection of DENV-4. Additionally, these commercial tests had decreased sensitivity in detecting secondary dengue infections, common in dengue endemic countries [3][4][5][6][7][8][9]. Commercially available NS1 antigen (Ag) tests manufactured by BioRad (Platelia) and Panbio (Dengue Early) both have low sensitivity to DENV-4 infections [8,[10][11][12].…”

Section: Introductionmentioning

confidence: 99%

“…Commercially available NS1 antigen (Ag) tests manufactured by BioRad (Platelia) and Panbio (Dengue Early) both have low sensitivity to DENV-4 infections [8,[10][11][12]. Previous evaluations of Dengue Early test showed a sensitivity as low as 19% for DENV4 [6,8,[13][14][15]. Although the overall sensitivity of Platelia test was good however when comparing the sensitivities between all DENV serotypes, the BioRad test had the lowest sensitivity for DENV-4 (58.3%) [10].…”

Section: Introductionmentioning

confidence: 99%

Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1)

Gelanew¹,

Hunsperger²

2018

Virol J

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BackgroundDengue, caused by one of the four serologically distinct dengue viruses (DENV-1 to − 4), is a mosquito-borne disease of serious global health significance. Reliable and cost-effective diagnostic tests, along with effective vaccines and vector-control strategies, are highly required to reduce dengue morbidity and mortality. Evaluation studies revealed that many commercially available NS1 antigen (Ag) tests have limited sensitivity to DENV-4 serotype compared to the other three serotypes. These studies indicated the need for development of new NS1 Ag detection test with improved sensitivity to DENV-4. An NS1 capture enzyme linked immunoassay (ELISA) specific to DENV-4 may improve the detection of DENV-4 cases worldwide. In addition, a serotype-specific NS1 Ag test identifies both DENV and the infecting serotype.MethodsIn this study, we used a small-ubiquitin-like modifier (SUMO*) cloning vector to express a SUMO*-DENV-4 rNS1 fusion protein to develop NS1 DENV-4 specific monoclonal antibodies (MAbs). These newly developed MAbs were then optimized for use in an anti-NS1 DENV-4 capture ELISA. The serotype specificity and sensitivity of this ELISA was evaluated using (i) supernatants from DENV (1–4)-infected Vero cell cultures, (ii) rNS1s from all the four DENV (1–4) and, (iii) rNS1s of related flaviviruses (yellow fever virus; YFV and West Nile virus; WNV).ResultsFrom the evaluation studies of the newly developed MAbs, we identified three DENV-4 specific anti-NS1 MAbs: 3H7A9, 8A6F2 and 6D4B10. Two of these MAbs were optimal for use in a DENV-4 serotype-specific NS1 capture ELISA: MAb 8A6F2 as the capture antibody and 6D4B10 as a detection antibody.ConclusionThis ELISA was sensitive and specific to DENV-4 with no cross-reactivity to other three DENV (1–3) serotypes and other heterologous flaviviruses. Taken together these data indicated that our MAbs are useful reagents for the development of DENV-4 immunodiagnostic tests.Electronic supplementary materialThe online version of this article (doi: 10.1186/s12985-018-0925-7) contains supplementary material, which is available to authorized users.

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Performance of commercial dengue NS1 ELISA and molecular analysis of NS1 gene of dengue viruses obtained during surveillance in Indonesia

Cited by 43 publications

References 40 publications

Clinical and virological characteristics of dengue in Surabaya, Indonesia

Clinical and virological characteristics of dengue in Surabaya, Indonesia

Detecting Dengue Virus Nonstructural Protein 1 (NS1) in Urine Samples Using ELISA for the Diagnosis of Dengue Virus Infection

Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1)

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