1. Immuno-reactivity against giants viruses by elisa sandwich to detect antibody versus elisa used nanobody to detect antigens in tunisian patients with pneumonia
Authors : Hanene Saadi, Claude Nappez, Mondher Boughalmi
Pages : 1-9
DOI : http://dx.doi.org/10.21172/1.133.01
Keywords : Mimiviridae, Megaviridae, Marseilleviridae, Zamilon virophage, ELISA, Nanobody, Pneumonia. Abstract :Several studies have used serological tests to assess the prevalence of Mimivirus infections in patients with pneumonia. And better study the implication of these viruses during this pathology. The results observed by several teams were contradictory between those who support the idea of involvement of giant amoeba viruses in this pathology and those who predict the absence of involvement during this human disease. To report more evidence, we wanted to test the sera of patients hospitalized for this pathology extemporaneously. For this, we have developed a high-throughput semi-automatic ELISA system to test the reactivity of serum antibodies "ELISA Acs" but also for the first time an ELISA based on Nanobody "ELISA Vhh" which tests the reactivity of Vhh against particles or antigens of giant amoeba viruses belonging to the families Mimiviridae, Megaviridae, Marseilleviridae and Zamilon. The use of viruses of Tunisian origin aims to determine the relationship between the ecological niche and the specific pathogens because of the important polymorphism and the significant chimerization of its viruses which has not been studied in the various serological reports. or based solely on the Mimivirus prototype. Preliminary results show a reactivity of Tunisian sera with Tunisian viruses by ELISA Acs, but it was also higher with the LBA111 virus belonging to the Megaviridae family than with any other virus tested. ELISA Vhh tests were negative with all sera from blood donors and patients. But positive with synthetic serum from repeated dose injection of each virus in laboratory rabbits and harvest after 48h. The Elisa tests with all the synthetic sera of the other viruses were conclusive with a remarkable positivity which makes it possible to validate the Vhh constructs and also the antibody production reaction by the rabbits. We propose protocol to standardize the ELISA studies for its giant viruses and compare the results on a non-random basis by fixing the way of calculation of cut-off and thus give a mathematical approach for the determination of positive or negative serum. our purpose and to give a standard ELISA protocol for testing its viruses and we also propose Vhh constructs directed against a panel of its viruses for high flow tests of sputum, nasopharyngeal fluid, broncho-alveolar lavage, bronchic aspiration and sera for sale online via our portal. It was also noted that the reproducibility of the results and the number of false negatives were more conclusive with the use of Vhh than with ELISA Acs.
Citing this Journal Article :Hanene Saadi, Claude Nappez, Mondher Boughalmi, "Immuno-reactivity against giants viruses by elisa sandwich to detect antibody versus elisa used nanobody to detect antigens in tunisian patients with pneumonia", Volume 13 Issue 3 - May 2019, 1-9
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