RESUMEN
Identifying the time of ovulation is an important process for women seeking and avoiding pregnancy. Luteinizing hormone (LH) plays an important role in ovulation, which is very important in the reproductive mechanism. Therefore, detecting the LH level is of great importance in monitoring ovulation. In this study, sensitive, rapid and selective electrochemical biosensors were developed to detect LH quantitatively from human urine samples and to monitor the ovulation period. Isopotential region and current density optimization studies revealed that sensors with an electrode width and spacing of 1 mm had the optimum performance. Electrochemical impedance spectra evidenced immobilization of DSP self-assembled monolayers and anti-LH-beta antibody on the surface. While the mobile phone vibrator led to a 3.5-fold enhancement in response signals, the agitation system developed resulted in a 10-fold improvement. The sensors displayed detection limits of 1.02 and 1.53 mIU/ml in the range of 0-40 mIU/ml LH concentration obtained using two statistical approaches. Additionally, the sensors showed no cross-reactivity to hCG, which is very similar in structure and is widely reported to have high cross-reactivity.
Asunto(s)
Técnicas Biosensibles , Hormona Luteinizante , Humanos , Hormona Luteinizante/orina , Femenino , Técnicas Biosensibles/métodos , Ovulación , Límite de Detección , Técnicas Electroquímicas , Electrodos , Técnicas Analíticas Microfluídicas/instrumentaciónRESUMEN
Human chorionic gonadotropin (hCG), a glycoprotein hormone secreted from the placenta, is an important biomarker for pregnancy. In this study, we designed a precise, rapid and fully automatic device with an electrochemical point-of-care biosensor capable of quantitative hCG detection from human urine samples for early pregnancy detection. Gold and Ag/AgCl electrodes, whose structure with optimum isopotential region and current density, were simulated using COMSOL Multiphysics® software and custom-made from Flex Medical. The sensing surface was fabricated with DSP self-assembled monolayers (SAMs) and covalently immobilized anti-hCG-beta antibody. The detection method involved a sandwich assay using anti-hCG alpha-HRP. Based on an automated agitation design implemented in our device, the surface reaction rate is significantly improved comparing to routinely performed sandwich assays, and therefore a rapid detection of very low concentration can be achieved. Electrochemical impedance spectroscopy (EIS) and chronoamperometry (CA) measurements were used to characterize the immobilization of the antibodies and to determine the sensor activities respectively. The sensors displayed a limit of detection (LOD) of 2.17 mIU/ml within established clinical hCG levels for early detection of pregnancy. They responded very well to hCG, but not to luteinizing hormone (LH), which has a high degree of cross-reactivity with hCG. The results showed that the immunosensor has high specificity, good reproducibility, and long-term stability for the detection of hCG in urine samples.