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In-Vitro Diagnostic Devices: Introduction to Current Point-of-Care Diagnostic Devices
Author:
ChaoMin Cheng / ChenMeng Kuan / ChienFu Chen
Publisher:
Springer
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Books / Home and family
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2015
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Addressing the origin, current status, and future development of point-of-care diagnostics, and serving to integrate knowledge and tools from Analytical Chemistry, Bioengineering, Biomaterials, and Nanotechnology, this book focusses on addressing the collective and combined needs of industry and academia (including medical schools) to effectively conduct interdisciplinary research.

In addition to summarizing and detailing developed diagnostic devices, this book will attempt to point out the possible future trends of development for point-of-care diagnostics using both scientifically based research and practical engineering needs with the aim to help novices comprehensively understand the development of point-of-care diagnostics. This includes demonstrating several common but critical principles and mechanisms used in point-of-care diagnostics that address practical needs (e.g., disease or healthcare monitoring) using two well-developed examples so far: 1) blood glucose meters (via electrochemistry); and, 2) pregnancy tests (via lateral flow assay).

Readers of this book will come to fully comprehend how to develop point-of-care diagnostics devices, and will be inspired to contribute to a critical global cause the development of inexpensive, effective, and portable in vitro diagnostics tools (for any purpose) that can be used either at home or in resource limited areas.

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Springer International Publishing Switzerland 2016

Chao-Min Cheng , Chen-Meng Kuan and Chien-Fu Chen In-Vitro Diagnostic Devices 10.1007/978-3-319-19737-1_1

1. Introduction to In Vitro Diagnostic Devices

Chao-Min Cheng 1

(1)

National Tsing Hua University, Hsinchu, Taiwan

(2)

National Chung Hsing University, Taichung, Taiwan

Chao-Min Cheng (Corresponding author)

Email:

Chen-Meng Kuan

Email:

Chien-Fu Chen

Email:

1.1 Overview

Healthcare investment keeps on increasing substantially in recent years [].

Molecular-based diagnostics is critical for prevention, identification, and treatment of disease. Current laboratory analyses support correct diagnosis in over 70 % of all diseases and can be used to aid the continuous monitoring of drug therapy [].

In addition to the improved efficiency in laboratory diagnostics, there has been a trend toward a more decentralized diagnostics which occurs directly at patients bedside, in outpatient clinics, or at the sites of accidents, so-called point-of-care (POC) systems [].

The first POC device was urine dipstick test, which was developed in 1957 to measure urinary protein []. Glucose meters for diabetic monitoring and lateral-flow devices for pregnancy tests are currently the most widely used devices in POC molecular diagnostics. They are excellent examples of POC tests; however, they are still not applicable if highly sensitive and high-throughput quantitative measurements are required.

In recent decades, some technologies have emerged that fulfill these requirements. Lateral-flow immunoassay (LFIA) devices, for example, which were originally proposed in the 1980s, remain popular largely because of their design simplicity.

Plotz and Singer invented the latex agglutination assay in 1956, from which the technical basis for the LFIA was derived []. Since that time, at least another 500 patents have been filed on various aspects of the technology. Several patents have even been formatted by companies such as Becton Dickinson & Co. and Unilever and Carter Wallace.

The chief application driving the early development of solid-phase, rapid-test technology was the human pregnancy test, which was symbolic of continued historical interest in urine testing for medical diagnostic purposes. This particular testing application made great strides in the 1970s, as a result of improvements in antibody generation technologies and significant gains in understanding the biology and detection of human chorionic gonadotropin (hCG), derived largely from the work performed by Vaitukaitis and colleagues []. However, to entirely evolve the lateral-flow test, considerable enabling technologies were still required. Many of these technologies, such as nitrocellulose membrane manufacturing, antibody generation, and processing equipment, were developed throughout the 1990s. The purpose of this article is to introduce readers with basic information regarding the LFIA approach.

1.2 Structure

Figure displays the key elements of a LFIA. This assay consists of several components, often segmented parts made of different materials. When a test is run, appropriately conditioned sample is added to the proximal end of the strip, the absorbent pad. The treated sample then migrates to the conjugate pad, where an appropriate reagent has been immobilized. The labeled reagent on the conjugate pad can be colloidal gold, or a colored, fluorescent, or paramagnetic latex particle. These specific biological components can be either antigen or antibody depending on the assay format. Next, the sample remobilizes the dried reagent, and particle interaction ensues. Sample and reagent then migrate to the next segment of the strip, the reaction matrix. The reaction matrix is a porous membrane, upon which a final specific biological component has been immobilized. These biological components are usually proteins, either antibody or antigen. They have been bound onto the specific lines of the membrane being used. As the sample and reagent reach this line, they are captured by the applied proteins, and excess liquid moves past this point and is taken up by the absorbent pad. The result is the detectable absence or presence of the test line, read by eyes or by other instruments.

Fig. 1.1

Typical structure of a LFIA strip

The LFIA may be of two different types: (1) direct (sandwich, Fig. b). Both types can accommodate qualitative, semiquantitative, and fully quantitative determinations. Direct assay is usually used when testing for larger analytes with multiple antigenic sites, such as hCG, dengue antigen, or human immunodeficiency virus (HIV). A positive result is indicated by the presence of a test line. The conjugated particles also reach and are captured at the control line. The control line typically comprises a speciesspecific anti-immunoglobulin antibody, specific for the antibody in the conjugate pad. Competitive assay is usually used when testing for small molecules with single antigenic determinants that cannot bind to antibodies on a test line simultaneously. In such cases, a positive result is indicated by the absence of a test line, but a control line may still form.

Fig. 1.2

a Direct solid-phase immunoassay. b Competitive solid-phase immunoassay

1.3 Advantages

LFIAs represent a well-established and very appropriate technology when applied to a wide variety of in vitro diagnostics (IVD) or field-use applications. The advantages of the LFIA are well known:

Technology is mature.

Manufacture is relatively easy: Equipment and processes are already developed and available.

They can be scalable to high-volume production.

They can be stored for 1224 months, often without refrigeration.

They are easy to use, requiring minimal operator-dependent steps and interpretation.

They can handle small volumes of multiple sample types.

They can be integrated with onboard electronics, reader systems, and information systems.

They have high sensitivity, specificity, and good stability.

Development and approval are relatively low cost and require a short timeline.

They are already present and accepted by the market: Minimal education is required for users and regulators.

1.4 Antibody

Although the physical components of the lateral-flow test strip and construction techniques play a major role, the most critical part of the LFIA is the appropriate antibody to provide antigen recognition. If we chose inappropriate antibody, it would not have ability to recognize the target antigen. Much time is spent determining the most suitable antibody for specific assays. Many scientists have spent a great deal of time figuring out the suitable antibodies to fit the assay.

The LFIA is particularly demanding in terms of the mass of the reagent used to drive the antibody and antigen interaction. When an antibody is used in a sandwich-type assay, they are applied at a ratio of 13 g per cm across the width of the nitrocellulose strip, in a line 1 mm wide and with a relatively shallow bed volume of 0.13 mm. This results in an antibody concentration of 1030 g per square cm, which is 25100 times that used in an enzyme-linked immunosorbent assay (ELISA), which can typically require a maximum concentration of 300 ng per square cm [].

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