Immunosensor for Detection of Inflammation in Amniotic Fluid

Preterm Premature Rupture of Membranes (PPROM) is a pregnancy complication. In this condition, the sac (amniotic membrane) surrounding the fetus breaks (ruptures) before week 37 of pregnancy. Once the sac breaks, pregnant woman has increased risk for infection. PPROM complicates 3 – 4% of all pregnancies and is up to 1/3 complicated by microbial invasion of the amniotic cavity (MIAC) leading to infection in amniotic fluid (AF) and development of intra-amniotic inflammation. Although this complication is usually asymptomatic, it is a major cause of preterm birth and neonatal morbidity and mortality worldwide. Neonates from these pregnancies are at increased risk of developing neonatal sepsis and impaired psychomotor development and other sometimes lifelong health consequences.

Our invention is point-of-care test (POCT) based on electrochemical immunosensor for simultaneous detection of three inflammatory protein biomarkers with high predictive value in AF collected by amniocentesis. The aim of the test is to confirm intra amniotic inflammation. The entire biosensor consists of an immunosorbent magnetically active microparticles modifed with specific antibody. After capturing the protein biomarker by the immunosorbent, the bioconjugate (IgG antibody conjugated with an electroactive indicator, specifically quantum dots or gold nanoparticles) is added. A final measurable signal proportional to the concentration of the inflammatory protein biomarker is provided by metal ions released from the electroactive indicator. Disposable screen printed three electrode sensors are used for simultaneous electrochemical analysis. Clinical relevance of selected protein biomarkers was determined based on the results of proteomic and antibody studies of AF of pregnant women with PPROM.

Advantages
POCT enables to speed up the process of confirmation or elimination inflammation in AF and can be easily performed and evaluated in minutes beside the patient´s bed and thus enables personalised approach to therapeutic intervention of the pregnant woman. Currently, determining the MIAC is not specific enough and is very time consuming and technically demanding. It is based on combination of cultivation and molecular methods. Further research is focused on test verification in cervicovaginal fluid to avoid amniocentesis.