A Stopped Flow System with Hydrodynamic Injection for Red Blood Cells Osmotic Fragility Test: Possibility for Automatic Screening of Beta-thalassemia Trait

Abstract

Simple instrumentation and procedure with stopped flow system coupled to hydrodynamic injection and spectrophotometric detection were developed for automatic osmotic fragility test (OFT). OFT is a test for abnormal red blood cells based on the kinetics of their rupturing in a hypotonic saline solution. A portion of red cells was merged on-line with a hypotonic saline solution. They were mixed while flowing into the detector and were stopped for a short period for continuously monitoring the change of turbidity based on a transmission signal. In this work, a possible application of the system for screening of beta-thalassemia trait is demonstrated. Descriptions of instrumentation development and parameter optimization are presented. The system offers advantages over the conventional batch-wise OFT in terms of automation, precision, analysis time and sample volume.

K-means Clustering and Hierarchical Cluster Analysis Coupled with Linear Discriminant Analysis to Classify Signals in Osmotic Fragility Test for Thalassemia Screening

“…OFT signals used in this paper were obtained from prior research (Khonyoung et al, 2009). From the dataset, 73 blood samples were defined, according to the hospital records, as 21 and 52 samples with positive and negative test defined as RT and RN in Table 1, respectively.…”

“…The surface areas of red blood cells of a thalassemia patient (positive tests) are larger than in a normal person (negative tests), so the blood cells are more likely to breakdown faster than in negative tests. OFT, coupled with stopped flow injection, was developed as an automated thalassemia screening tool that reduce the risk of contamination in the system (Khonyoung et al, 2009). The signals of stopped flow OFT are used to discriminate patients.…”

“…The signals of stopped flow OFT are used to discriminate patients. The method can classify patients by using the optimized slope from the optimization based on hospital records (Khonyoung et al, 2009). However, no evidence exists guaranteeing its accuracy in predicting unknown samples.…”

Flow Injection/Sequential Injection Analysis Systems: Potential Use as Tools for Rapid Liver Diseases Biomarker Study

“…Some downscaled batch methods are able to reduce the volume to μ L, but in general, FI usually requires relatively less sample volume for a particular analyte or sample being studied. For example, the osmotic fragility test (OFT) of red blood cells normally requires 20  μ L of undiluted blood sample in batch spectrometric method whereas only 1  μ L of undiluted sample is required in the FI system where it is tested in 100-fold dilution [25]. As compared to standard bioassay technique such as ELISA, the volume required by flow-based systems is also usually lower.…”

Flow Injection Analysis System Using Magnetic Beads, Screen Printed Electrodes and Magnets

“…The concept of FIA was first discussed by Ruzicka and Hansen 1 and then the Stewart group 2 in the middle of 1970s. FIA offers other advantages as simple configuration, flexibility, adaptability to various chemistries and are classified into the basic mode (also known as normal flow injection analysis) and a wide variety of special modes, for example, stopped flow 3, 4 and reverse flow 5, 6. Since parts of the systems can be homemade or replaced as suitable, the cost of the systems is relatively lower than many other commercial instrumentation‐based analysis systems.…”