BackgroundEarly diagnosis of dengue can assist patient triage and management and prevent unnecessary treatments and interventions. Commercially available assays that detect the dengue virus protein NS1 in the plasma/serum of patients offers the possibility of early and rapid diagnosis.Methodology/Principal FindingsThe sensitivity and specificity of the Pan-E Dengue Early ELISA and the Platelia™ Dengue NS1 Ag assays were compared against a reference diagnosis in 1385 patients in 6 countries in Asia and the Americas. Platelia was more sensitive (66%) than Pan-E (52%) in confirmed dengue cases. Sensitivity varied by geographic region, with both assays generally being more sensitive in patients from SE Asia than the Americas. Both kits were more sensitive for specimens collected within the first few days of illness onset relative to later time points. Pan-E and Platelia were both 100% specific in febrile patients without evidence of acute dengue. In patients with other confirmed diagnoses and healthy blood donors, Platelia was more specific (100%) than Pan-E (90%). For Platelia, when either the NS1 test or the IgM test on the acute sample was positive, the sensitivity versus the reference result was 82% in samples collected in the first four days of fever. NS1 sensitivity was not associated to disease severity (DF or DHF) in the Platelia test, whereas a trend for higher sensitivity in DHF cases was seen in the Pan-E test (however combined with lower overall sensitivity).Conclusions/SignificanceCollectively, this multi-country study suggests that the best performing NS1 assay (Platelia) had moderate sensitivity (median 64%, range 34–76%) and high specificity (100%) for the diagnosis of dengue. The poor sensitivity of the evaluated assays in some geographical regions suggests further assessments are needed. The combination of NS1 and IgM detection in samples collected in the first few days of fever increased the overall dengue diagnostic sensitivity.
Background Dengue virus (DENV) infection remains a major public health burden worldwide. Soluble mediators may play a critical role in the pathogenesis of acute DENV infection. Galectin-9 (Gal-9) is a soluble β-galactoside-binding lectin, with multiple immunoregulatory and inflammatory properties. Objective To investigate plasma Gal-9 levels as a biomarker for DENV infection. Study design We enrolled 65 DENV infected patients during the 2010 epidemic in the Philippines and measured their plasma Gal-9 and cytokine/chemokine levels, DENV genotypes, and copy number during the critical and recovery phases of illness. Results During the critical phase, Gal-9 levels were significantly higher in DENV infected patients compared to healthy or those with non-dengue febrile illness. The highest Gal-9 levels were observed in dengue hemorrhagic fever (DHF) patients (DHF: 2464 pg/ml; dengue fever patients (DF): 1407 pg/ml; non-dengue febrile illness: 616 pg/ml; healthy: 196 pg/ml). In the recovery phase, Gal-9 levels significantly declined from peak levels in DF and DHF patients. Gal-9 levels tracked viral load, and were associated with multiple cytokines and chemokines (IL-1α, IL-8, IP-10, and VEGF), including monocyte frequencies and hematologic variables of coagulation. Further discriminant analyses showed that eotaxin, Gal-9, IFN-α2, and MCP-1 could detect 92% of DHF and 79.3% of DF, specifically (P<0.01). Conclusion Gal-9 appears to track DENV inflammatory responses, and therefore, it could serve as an important novel biomarker of acute DENV infection and disease severity.
The prevalence study and the characterization of hepatitis C virus (HCV) have been carried out in the Philippines and the sequence determination of 5'-untranslated region (5'-UTR)-Core and NS5B regions of HCV was performed in this study. An HCV strain (SE-03-07-1689) collected in Metro Manila, Philippines, belonged to discordant subtypes, 2b and 1b in 5'-UTR-Core and NS5B regions, respectively. The 9.3kb-sequence of this strain including the entire open reading frame were compared with those of the reference strains retrieved from the HCV sequences database (GenBank/EMBL/DDBJ) and indicated a recombination event.The computation of the sequence similarity mapped a crossover point within NS3region. This is the second report on the inter-genotype recombinant of HCV and the third one when an intra-genotype recombinant is included. This recombinant strain, SE-03-07-1689, is tentatively designated as RF3_2b/1b according to the suggestions used for other two HCV recombinants.
In order to prevent cervical cancer, vaccines against human papilloma virus types 16 (HPV-16) and 18 (HPV-18) have been implemented worldwide. However, the HPV types that cause cancer can differ according to geographical area and ethnicity. In this new era of the HPV vaccine, it is important to elucidate the prevalent HPV types in each area. Therefore, the prevalence of HPV infection and cervical abnormalities among 369 female commercial sex workers in the Philippines were examined. HPV L1 gene was amplified by polymerase chain reaction (PCR) using modified GP5+/6+ primers, and genotyping was performed by sequencing cloned PCR products. HPV DNA was detected in 211 (57.2%) women, among whom 46 HPV types were identified. HPV-52 was most common and multiple-type infection was observed in 44.5%. Among 56 women with abnormal cervical cytology (low- and high-grade squamous intraepithelial lesions and adenocarcinoma in situ), HPV-52 was most common (23.2%), followed by HPV-16 (19.6%), -58 (10.7%), and -67 (10.7%). Only 27% of these women were positive for HPV-16 and -18. Multivariate analysis revealed that HPV-16, -39, -52, -67, and -82 were significantly associated with abnormal cytology. Repeated analysis of HPV-52 single-positive samples using the original GP5+/6+ PCR primers produced negative results in 57% of cases, suggesting that the prevalence of HPV-52 infection may have been underestimated in previous studies, and the current vaccines may not be sufficient for preventing infection and the development of premalignant lesions of the cervix in women in the Philippines.
Introduction Dengue virus (DENV) is transmitted by the mosquito vector, and causes a wide range of symptoms that lead to dengue fever (DF) or life-threatening dengue hemorrhagic fever (DHF). The host and viral correlates that contribute to DF and DHF are complex and poorly understood, but appear to be linked to inflammation and impaired coagulation. Full-length osteopontin (FL-OPN), a glycoprotein, and its activated thrombin-cleaved product, trOPN, integrate multiple immunological signals through the induction of pro-inflammatory cytokines. Materials and Method To understand the role of OPN in DENV-infection, we assessed circulating levels of FL-OPN, trOPN, and several coagulation markers (D-dimer, thrombin-antithrombin complex [TAT], thrombomodulin [TM], and ferritin in blood obtained from 65 DENV infected patients in the critical and recovery phases of DF and DHF during a dengue virus epidemic in the Philippines in 2010. Results Levels of FL-OPN, trOPN, D-dimer, TAT, and TM were significantly elevated in the critical phase in both the DF and DHF groups, as compared with healthy controls. During the recovery phase, FL-OPN levels declined while trOPN levels increased dramatically in both the DF and DHF groups. FL-OPN levels were directly correlated with D-dimer and ferritin levels, while the generation of trOPN was associated with TAT levels, platelet counts, and viral RNA load. Conclusion Our study demonstrated the marked elevation of plasma levels of FL-OPN and thrombin-cleaved OPN product, trOPN, in DENV-infection for the first time. Further studies on the biological functions of these matricellular proteins in DENV-infection would clarify its pathogenesis.
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