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A Microfluidic Chip-Based MRS Immunosensor for Biomarker Detection via Enzyme-Mediated Nanoparticle Assembly.

ABSTRACT: Conventional immunoassay methods have their common defects, such as tedious processing steps and inadequate sensitivity, in detecting whole blood. To overcome the above problems, we report a microfluidic chip-based magnetic relaxation switching (MRS) immunosensor via enzyme-mediated nanoparticles to simplify operation and amplify the signal in detecting whole blood samples. In the silver mirror reaction with catalase (CAT) as the catalyst, H₂O₂ can effectively control the production of Ag NPs. The amount of Ag NPs formed further affects the degree of aggregation of magnetic nanoparticles (MNP_S), which gives rise to the changes of transverse relaxation time (T₂). Both sample addition and reagent reaction are carried out in the microfluidic chip, thereby saving time and reagent consumption. We also successfully apply the sensor to detect alpha-fetoprotein (AFP) in real samples with a satisfied limit of detection (LOD = 0.56 ng/ml), which is superior to the conventional ELISA.

SUBMITTER: Yin B

PROVIDER: S-EPMC8193930 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The rapid and accurate diagnostic methods to identify carbapenemase-producing organisms (CPO) is of great importance for controlling the CPO infection. Herein, we have developed a microfluidic chip-based technique to detect CPO and assessed its clinical value in detecting CPO directly from blood cultures (BCs). The detection performance of the microfluidic chip-based LAMP amplification method was analyzed retrospectively on a collection of 192 isolates including molecularly characterized 108 CPO and 84 non-CPO and prospectively on a collection of 133 positive BCs with or without CPO suspicion, respectively. In the retrospective study, the microfluidic chip-based LAMP amplification method exhibited 87.5% accuracy (95% CI [82.0-91.5]), 97.7% sensitivity (95% CI [91.2-99.6]), 78.8% specificity (95% CI [69.5-86.0]), 79.6% positive predictive value (PPV) (95% CI [70.6-86.5]) and 97.6% negative predictive value (NPV) (95% CI [90.9-99.6]). Among the 192 isolates, 22 (11.5%) false-positives (FP) and 2 (1.0%) false negatives (FN) were observed. In the prospective study, the 133 routine isolates of positive BCs including 18 meropenem-resistant CPO and 115 non-CPO were assessed, and 4 FP were observed in non-CPO and CPO, respectively. The current method showed a total detection performance of 94.0% accuracy (95% CI [88.4-97.1]), 100.0% sensitivity (95% CI [73.2-100.0]), 93.2% specificity (95% CI [86.7-96.8]), 63.6% PPV (95% CI [40.8-82.0]) and 100.0% NPV (95% CI [95.8-100.0]). In summary, the microfluidic chip-based LAMP amplification method is reliable for the rapid screening and detection of CPO with high accuracy, sensitivity, and specificity, and could easily be implemented in clinical microbiology laboratories. IMPORTANCE Rapid and accurate identification of CPO may reduce the genetic exchanges among bacteria and prevent further dissemination of carbapenemases to non-CPO. The current method had designed microfluidic chip-based LAMP amplification method for multiplex detection of carbapenemase genes and evaluated the detection performance of the newly method. The current method can rapidly screen and detect CPO with high accuracy, sensitivity, and specificity, and could easily be implemented in clinical microbiology laboratories, as this will reduce the carbapenem resistance issues worldwide.

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A Microfluidic Chip-Based MRS Immunosensor for Biomarker Detection via Enzyme-Mediated Nanoparticle Assembly.

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