Ten years since the introduction of bacterial glycoengineering technology in the diagnostic algorithm of Hemolytic Uremic Syndrome

LUCIANO J. MELLI; STELLA MARIS LANDIVAR; MILIWEBSKY, ELIZABETH; ISABEL CHINEN; DIEGO J. COMERCI; JUAN E. UGALDE; ANDRÉS E. CIOCCHINI

Rosario

Congreso; LIX Congreso Anual de la Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular (SAIB 2023); 2023

SAIB

Human infection with Shiga toxin-producing Escherichia coli (STEC) is a major cause of postdiarrheal hemolytic uremic syndrome (HUS), a life-threatening condition characterized by hemolytic anemia, thrombocytopenia and acute renal failure. E. coli O157:H7 is the dominant STEC serotype associated with HUS worldwide although non-O157 STEC serogroups can cause a similar disease. In Argentina, O157, O145, O121, and O103 are the most prevalent serogroups. Detection of anti-O157 LPS antibodies in combination with bacteriological procedures considerably improves diagnosis of STEC infections. Ten years ago, we have exploited the bacterial glycoengineering technology for the development of recombinant glycoproteins consisting of the O157, O145, O121 or O103-polysaccharide attached to a carrier protein. Then, an indirect ELISA was developed by using these serogroup-specific bacterial engineered glycoproteins (Glyco-iELISAs). We demonstrate that using these antigens it is possible to clearly discriminate between STEC O157, O145 and O121 infected patients and healthy children, even at early stages of the disease, as well as to confirm the diagnosis in HUS patients in which the classical diagnostic procedures failed. The Glyco-iELISAs were transferred to the National Reference Laboratory (NRL) where more than 1800 samples were analyzed over a period of 10 years. The implementation of Glyco-iELISAs in the NRL allowed to increase the diagnosis of association to HUS to STEC from 25 to 75%. In recent years, advances in the field of nanobiotechnology and their application for the development of lateral flow immunoassays (LFIA) took on great relevance as tests for the rapidand in place diagnosis of different infectious diseases. Using the LFIA platform, we developed CHEMSTRIP® E. coli O157/O145 test that allows the detection of specific anti-O157 and anti-O145 IgM antibodies using the recombinant glycoproteins AcrA-O157 and AcrA-O145, respectively. By analyzing sera from patients with information from the days post-onset of symptoms, it was possible to detect 100% of positive cases after 3 days of onset of symptoms and up to 80% of positive cases before 3 days. The intervention of these antigens in the diagnostic algorithm produced a significant improvement in the association of HUS / DS to STEC and we consider that the implementation of the newly developed test could become an effective tool to avoid delays in supportive treatment or the application of a specific treatment when this becomes available.