Escherichia coli Enumeration in a Capillary-Driven Microfluidic Chip with SERS.

Pathogen detection is still a challenging issue for public health, especially in food products. A selective preconcentration step is also necessary if the target pathogen concentration is very low or if the sample volume is limited in the analysis. Plate counting (24-48 h) methods should be replaced by novel biosensor systems as an alternative reliable pathogen detection technique. The usage of a capillary-driven microfluidic chip is an alternative method for pathogen detection, with the combination of surface-enhanced Raman scattering (SERS) measurements. Here, we constructed microchambers with capillary microchannels to provide nanoparticle-pathogen transportation from one chamber to the other. Escherichia coli (E. coli) was selected as a model pathogen and specific antibody-modified magnetic nanoparticles (MNPs) as a capture probe in a complex milk matrix. MNPs that captured E. coli were transferred in a capillary-driven microfluidic chip consisting of four chambers, and 4-aminothiophenol (4-ATP)-labelled gold nanorods (Au NRs) were used as the Raman probe in the capillary-driven microfluidic chip. The MNPs provided immunomagnetic (IMS) separation and preconcentration of analytes from the sample matrix and then, 4-ATP-labelled Au NRs provided an SERS response by forming sandwich immunoassay structures in the last chamber of the capillary-driven microfluidic chip. The developed SERS-based method could detect 101-107 cfu/mL of E. coli with the total analysis time of less than 60 min. Selectivity of the developed method was also tested by using Salmonella enteritidis (S. enteritidis) and Staphylococcus aureus (S. aureus) as analytes, and very weak signals were observed.

A capillary driven microfluidic chip for SERS based hCG detection.

In this study, a capillary driven microfluidic chip-based immunoassay was developed for the determination of Human Chorionic Gonadotropin (hCG) protein, which is prohibited by the World Anti-Doping Agency (WADA). Here, we used antibody modified magnetic metal organic framework nanoparticles (MMOFs) as a capture prob in urine sample. MMOF captured hCG was transferred in a capillary driven microfluidic chip consisting of four chambers, and the interaction of MMOF with gold nanorods labelled with 5,5'-Dithiobis-(2-nitrobenzoic acid) (DTNB) as a Raman label was carried out in the capillary driven microfluidic chip. The movement of MMOF through first chamber to the last chamber was achieved with a simple magnet. In the last chamber of capillary driven microfluidic chip, SERS signals of DTNB molecules from the sandwich complex were recorded using a Raman spectrophotometer. The selectivity of the developed method was demonstrated by applying the same procedure for the detection of Human Luteinizing Hormone (hLH), Human Chorionic Gonadotropin Hormone (hGH) and Immunoglobulin G (IgG) protein. The regression coefficient and limit of detection obtained from the standard addition method were found as 0,9985 and 0,61 IU/L, respectively. Furthermore, the conventional ELISA method confirmed that the results obtained by the presented method were acceptable with the similarity of 97.9% in terms of average recovery value, for the detection of hCG in urine samples. The analysis system developed for target proteins will be an alternative technique such as Western Blot used in routine analysis that is expensive and time consuming.

Multiplex enumeration of Escherichia coli and Salmonella enteritidis in a passive capillary microfluidic chip.

Multiplex detection and quantification of bacteria in water by using portable devices are particularly essential in low and middle-income countries where access to clean drinking water is limited. Addressing this crucial problem, we report a highly sensitive immunoassay sensor system utilizing the fluorescence technique with magnetic nanoparticles (MNPs) to separate target bacteria and two different types of quantum dots (CdTe and Ni doped CdTe QDs) incorporated into a passive microfluidic chip to transport and to form sandwich complexes for the detection of two target bacteria, namely Escherichia coli (E. coli) and Salmonella enteritidis (S. enteritidis) in less than 60 min. The assay is carried out on a capillary driven microfluidic chip that can be operated by merely pipetting the samples and reagents, and fluorescence measurements are done by using a handheld fluorescence spectrophotometer, which renders the system portable. The linear range of the method was found to be 101 to 105 cfu mL-1 for both E. coli and S. enteritidis. The limit of detection (LOD) was calculated to be 5 and 3 cfu mL-1 for E. coli and S. enteritidis, respectively. The selectivity of the method was examined by testing Enterobacter dissolvens (E. dissolvens) and Staphylococcus aureus (S. aureus) samples, and no significant interference was observed. The method was also demonstrated to detect bacteria in tap water and lake water samples spiked with target bacteria.