INQUISAL   20936
INSTITUTO DE QUIMICA DE SAN LUIS "DR. ROBERTO ANTONIO OLSINA"
Unidad Ejecutora - UE
artículos
Título:
Laser-induced fluorescence integrated in a microfluidic immunosensor for quantification of human serum IgG antibodies to Helicobacter pylori
Autor/es:
MARCO A. SEIA; SIRLEY V. PEREIRA; CARLOS A. FONTÁN; IRMA E. DE VITO; GERMÁN A. MESSINA; JULIO RABA
Revista:
SENSORS AND ACTUATORS B-CHEMICAL
Editorial:
ELSEVIER SCIENCE SA
Referencias:
Lugar: Amsterdam; Año: 2012 vol. 168 p. 297 - 302
ISSN:
0925-4005
Resumen:
A portable immunosensor coupled to laser-induced fluorescence (LIF) detection system has been successfully developed. It was applied to the quantitative determination of specific IgG antibodies against Helicobacter pylori present in human serum samples. This system is a relevant alternative tool for the diagnostic of peptic ulcer and gastritis diseases produced by H. pylori, which are recognized risk factors for the development of gastric mucosa associated lymphoid tissue lymphoma and gastric adenocarcinoma. More than 50% of the population has been affected by H. pylori, one of the most widespread infections around the world. Our proposed system combines the LIF detection, which offers great sensibility with the specificity of the immunological reactions and the microfluidic technology. The device has a central channel (CC) with packed H. pylori antigen immobilized on 3-aminopropyl-modified controlled pore glass (AP-CPG). Antibodies in serum samples reacted immunologically with the immobilized antigen and then, they were determined using alkaline phosphatase (AP) enzyme-labeled second antibodies specific to human IgG. The 4-methylumbelliferyl phosphate (4-MUP), employed as enzymatic substrate, was converted to soluble fluorescent methylumbelliferone by AP, and this fluorescent product was finally quantified by LIF detection system. The calculated detection limits for LIF detection and the ELISA procedure were 0.17 and 2.1 U mL -1, respectively, and the within- and between-assay coefficients of variation were below 5.1%. © 2012 Elsevier B.V.