Development of double antibody sandwich ELISA as potential diagnostic tool for rapid detection of Crimean-Congo hemorrhagic fever virus

Abstract

Crimean-Congo hemorrhagic fever (CCHF) virus, a highly pathogenic viral agent is responsible for severe fatal hemorrhagic infections in many parts of the world. The early diagnosis of CCHF infection is important for successful clinical management and epidemiological control. The nucleoprotein (NP) of CCHFV being highly conserved and immunogenic is used as early diagnostic marker. In this study, we report a rapid and sensitive double antibody based antigen capture ELISA to detect Crimean-Congo hemorrhagic fever virus (CCHFV). Highly specific polyclonal and monoclonal antibody against NP has been generated and used as capture and detector antibody respectively. The assay was able to detect viral nucleoprotein in different matrices including human serum, ticks and culture supernatant. The detection limit of the developed sandwich ELISA assay was 25 ng of purified antigen. Comparison with a real time RT-PCR revealed its detection limit to be 1000 genome equivalents of CCHFV. Further the assay was comparatively evaluated with a commercial kit employing gamma irradiated CCHFV, revealing a sensitivity and specificity of 100%. This newly developed sandwich ELISA (sELISA) with high sensitivity and specificity could be used as an efficient method for the detection of CCHF virus in humans, ticks and culture supernatant. The assay will be useful as alternate tool for diagnosis of acute infection and is amenable for screening of large scale samples in resource limited settings.

Leukotriene metabolism and proiflammatory cytokines in Crimean Congo hemorrhagic fever

“…The determination of the severe cases and mortality in the early stage is very important to arrangement of therapeutic strategy, epidemiological control and optimal use of resources such as intensive care units. 14 In the present study we did not find any differences between severe and mild-moderate groups in terms of 5-LO and CYSLT levels, but higher IL-6 and ferritin levels were found in severe group compared to the mild group. We also observed higher CYSLT, IL-6, and ferritin levels in survive group compared to F I G U R E 5 Evaluation of the diagnostic performance of the interleukin-6 and ferritin in discriminating diseases mortality nonsurvive group.…”

Development and Characterization of Phage-Display-Derived Novel Human Monoclonal Antibodies against the Receptor Binding Domain of SARS-CoV-2

“…Sandwich ELISA is a reliable and rapid detection tool for infectious viral antigens [ 35 , 36 ]. To establish a sandwich ELISA specific to wild-type SARS-CoV-2 RBD antigen, K102.1 was used as the capture antibody whereas K102.2-HA was used as the detection antibody.…”

Single Molecule Level and Label-Free Determination of Multibiomarkers with an Organic Field-Effect Transistor Platform in Early Cancer Diagnosis

“…For instance, the development of an accurate and reliable method, such as enzyme-linked immunosorbent assay (ELISA), has received significant attention in early cancer diagnosis. 1 However, the ELISA method usually needs a long time with professional testers as well as complicated experimental processes. Besides, a limited quantification range is not sufficient for the detection of ultralow abundance proteins.…”

“…After acidifying with concentrated hydrochloric acid, collecting by filtration, and washing with deionized water, the final product pillar [5]arene-COOH was dried under vacuum. 1 H NMR (400 MHz, DMSO) δ 13.26 (s, 4H), 8.44 (s, 2H), 7.94 (s, 4H), 7.27 (s, 2H), 6.70 (s, 2H), 6.63 (s, 2H), 6.25 (s, 2H), 5.79 (s, 2H), 3.69 (d, J = 41.9 Hz, 9H), 3.60−3.47 (m, 16H), 3.10 (s, 8H).…”

“…Scientific research for people’s health plays an irreplaceable role in our social life. For instance, the development of an accurate and reliable method, such as enzyme-linked immunosorbent assay (ELISA), has received significant attention in early cancer diagnosis . However, the ELISA method usually needs a long time with professional testers as well as complicated experimental processes.…”