Comparative assessment of commercial enzyme-linked immunosorbent assay & rapid diagnostic tests used for dengue diagnosis in India.

Background & objectives: Dengue diagnosis is routinely carried out by detection of dengue virus (DENV) antigen NS1 and/or anti-DENV IgM antibodies using enzyme-linked immunosorbent assays (ELISAs) and rapid diagnostic tests (RDTs). This study was aimed at evaluation of quality of diagnostic assays currently in use in India for the identification of DENV infection. Methods: During 2016 dengue season (July-November) in Pune, India, comparative assessment of a few immunoassays was undertaken using (i) WHO-approved Panbio-Dengue-Early-(NS1)-ELISA and Panbio-Dengue-IgM-Capture-ELISA as reference tests, and (ii) Bayesian latent class analysis (BLCA) which assumes that no test is perfect. The assays included J.Mitra-Dengue-NS1-Ag-MICROLISA (JME-NS1), J.Mitra-Dengue-IgM-MICROLISA (JME-IgM), and two RDTs, namely, J.Mitra-Dengue-Day-1-Test (JM-RDT) and SD-BIOLINE-Dengue-Duo (SDB-RDT). Serum samples from patients seeking dengue diagnosis (n=809) were tested using the diagnostic kits. The presence of NS1 and/or IgM was taken as evidence for dengue-positive diagnosis. Results: Panbio-NS1/IgM-ELISAs identified 38.6 per cent patients as dengue positive. With Panbio-ELISA as reference, all the tests were less sensitive for IgM detection, while for NS1, JM-RDT was less sensitive. For combined diagnosis (both markers), sensitivity of all the tests was low (55.7-76.6%). According to BLCA, Panbio-ELISA was 84 per cent sensitive for NS1, 86 per cent specific for IgM and 87 per cent specific for combined diagnosis. Accordingly, performance of the other tests was substantially improved with BLCA; however, sensitivity of both the RDTs for IgM detection remained unacceptable. The NS1 ELISAs and RDTs detected all four DENV serotypes, JME being most efficient. All IgM tests exhibited higher sensitivity in secondary infections. Interpretation & conclusions: These results confirmed superiority of ELISAs, and testing for both NS1 and IgM markers for dengue diagnosis, and emphasized on improvement in sensitivity of RDTs.

Risk of transfusion-associated dengue: screening of blood donors from Pune, western India.

BACKGROUND: Dengue, a mosquito-borne viral disease, is endemic in >125 countries worldwide. The threat of blood-borne transmission of dengue virus (DENV) has been documented. STUDY DESIGN AND METHODS: This study was conducted to assess the potential magnitude of transfusion-associated dengue, by determination of DENV seromarkers in blood donations from Pune, India, during two dengue seasons (2016 and 2017). These included DENV nonstructural protein 1 (NS1), anti-DENV immunoglobulin (Ig) M, anti-DENV IgG (enzyme-linked immunosorbent assay), and DENV RNA (reverse transcription-polymerase chain reaction). RESULTS: NS1 (IgM) reactivity was 1 of 209, 0.48% (11/209, 5.3%) in 2016 and 2 of 311, 0.64% (20/311, 6.4%) in 2017. Of the 34 NS1/IgM reactives, 1 NS1-reactive donor and 10 IgM-reactive donors exhibited evidence of secondary infection. DENV RNA was not detected in any of the 34 NS1/IgM reactives. Among the NS1/IgM negatives, anti-DENV IgG reactivity was high in 2016 (75%) and further increased in 2017 (87%, p = 0.002). CONCLUSION: Although RNA negative, detection of 34 NS1/IgM-reactive donations, of which 11 had evidence of secondary infection, suggests the need for further evaluation on the basis of potential risk to recipients of either dengue transmission or increased risk of secondary infection. These would include multicenter studies followed by cost-benefit analyses.