UNIVERSITY OF SOUTHAMPTON

Development of Lab On A Chip device for Point Of Care Testing

Lab On A Chip Coursework

Bishwojit Konsam

Development of Lab On A Chip device for Point Of Care Testing Introduction

Point of care (diagnostic) test (POCT) is a test performed at the location of the patient without the necessity of clinical lab. Lab on a chip (LOAC) technology being a technology whose characteristic is low consumption of sample and reagent, small size, mobile, efficient and provides fast accurate analysis results, makes it a perfect technology for POCT devices. This essay describes the development of LOAC device for point of care testing. It also outlines some of the benefits of the technology. It also discusses about the future trends of the technology as POCT device. It also discusses about the barrier associated with the application of the technology and its possible solutions.

Benefits of POCT device

POCT device provides rapid and accurate results which in turn can be used to monitor patients so that rapid intervention can be provided to the patient as required. It also provides solution to the ever expanding range of medical test [1, 2]. The POCT has the capability to improve and modernize healthcare and also improve clinical outcomes [3]. A comparison of common clinical blood testing and point of care testing is given in the table 1[4]. Table 1: Comparison of clinical blood testing and point-of-care testing Laboratory based system 1. Test is ordered 2. Test request is processed 3. Nurse draws blood sample 4. Sample is transported to the lab 5. Sample is labelled and stored 6. Sample is centrifuged 7. Serum sample is sorted to analysers 8. Sample is analysed 9. Results are reviewed by the lab staff 10. Results are reported to the department 11. Clinician acts on results Point-of-care system 1. 2. 3. 4. Test is ordered Nurse draws blood sample Sample is analysed Results are reviewed by a nurse 5. Clinician acts on results

Another benefit of POCT device over centralise clinical testing is the time it saves. Some of the frequently measured compounds and their time scale for monitoring is shown in table 2 [7]. As in the table some of the compounds take hours to days to monitor but with POCT device the time scale may be highly reduced.

Table 2: Monitoring time for some frequently measure compounds

Seconds/minutes O2 CO2 K+ Glucose Lactate Cortisol Neurotransmitters Hours Creatinine Bilirubin Urea Sodium Chloride Triglyceride Days Iron Albumin Globulin Cholesterol

Developments in lab on chip devices for point of care testing

The earliest point of care device was a glucose detection device based on tablets containing test reagents. POCT devices were further improved into a dip stick devices as a result of technical innovations. At present, most of the POCT devices are self contained and are based on lateral flow test which requires addition of samples only. LOAC technology has a versatile fluid handling and integrated detection capability. This makes LOAC a promising technology for future POCT device which will require assay of high technical complexity. As a POCT device, the LOAC system should be very easy to use, portable and efficient and also should provide reliable and accurate results. The clinical potential of microchip capillary electrophoresis was reviewed by Colyer et al. in 1997 [5]. Since then, the concept of LOAC has grow rapidly and has been applied to range of applications ranging from clinical diagnostics, through genomics and proteomics research, to high throughput screening and drug discovery. The first commercial LOAC product based upon capillary gel electrophoresis was developed by Caliper and Agilent in 1999. The following paragraphs describe the early development in LOAC devices for POCT device. An online blood electrolyte monitoring system based on flow-through cell in a sandwich arrangement was demonstrated by Gumbrecht et al in 1990 [6]. The device was made up of a chip carrier, a chem FET chip and a PMMA slide with an engraved flow through channel. It was possibly the earliest clinical LOAC device [7]. It was designed to monitor pH, K+ and Ca2+ ion concentration from a 10 l blood. Another device for monitoring K+ ion in blood was demonstrated by Uhlig et al. which was capable of operating for at least 95 hr with a signal variation of 2% in blood [8]. The measurement of blood gases and electrolytes was successfully performed by using a florescence detection system [9]. It could perform 2000 measurements without calibration. A system consisting of microdialysis sampling interface, a silicon microflow manifold and an integrated biosensor array was demonstrated to monitor glucose and lactate in real time [10, 11]. Another device based on microdialysis was developed to

monitor clinically relevant substances such as glucose, lactate and metal ions. It was developed by Bohm et al. [12]. It was made up of two modules, an integrated low volume microdialysis probe which was connected to channel system in silica. It was driven by an external pump. The first microchip based capillary electrophoresis system for immunoassay was developed to perform separation of the products of homogenous immunological reactions in 1997 [13]. Another integrated microsystem for immunoassay was presented by Attiya et al. which consisted of a sample introduction unit, mixing device, reaction chamber, separation and self calibration unit [14]. It was able to perform a direct immunoassay of fluorescein labelled bovine serum albumin. The first microfluid chip for heterogenous bioassay based on electrokinetic was demonstrated in 2001 [15]. It was based on a locally immobilized protein A layer for detection of fluorescently labelled rabbit IgG. A personal genetic diagnostic device was introduced by Mathias et al. which was used to determine X and Y chromosomes. It was based on a typical 200 bp segment which appeared in male samples while absent in female samples [16, 17]. Biosite inc. developed a microfluidic immunoassay device which was able to perform quantitative immunoassay. It consisted of a portable fluorimeter and a protein chip. It was developed to be used as an aid in diagnosis of myocardial infarction, congestive heart failure and drugs of abuse overdose. It could determine quantitative concentration by performing simultaneous immunoassay from whole blood using controlled fluid flow [18]. A device based on gel filled planar microstructure was developed to separate fluorescent labelled amino acids [19]. Another device based on micellar electrokinetic chromatography (MEKC) was developed for the separation of biological samples [20]. The separation was one to two order magnitudes faster than fused silica capillaries. A disposable PMMA microfluidic systems for high-throughput CE applications with an elastic PDMS cover was developed by Boone and Hooper [21]. A disposable ELISA for DDdimer was demonstrated by Schwartz et al. which could serve as a diagnostic aid in suspected venous thromboembolism in blood plasma [22]. Micronics Inc. developed a device called H-filter [23]. It is a hydrostatic pressure driven filter which could perform micro separation and extraction based on diffusion. It allows separation of small particles larger particles continuously without the need for filtration, centrifugation or dialysis. Micronics Inc. also developed self calibrating micro chemical reactor and disposable microfluidic system called T sensor [24, 25]. It is a simple T shaped device and it is used for direct optical detection and quantitation of human serum albumin in whole blood. Current developments An easy to handle and disposable clinical diagnostic device was developed using fully integrated plastic microfluidic components. It is capable to sample and identify

which make the measurements of metabolic parameters in human blood fast and accurately. The device could measure metabolic parameters like glucose, lactate and partial oxygen from human whole blood with the help of a portable analyzer [26]. A polymer based device for reverse transcription (RT) - polymerase chain reaction (PCR) was developed for POCT diagnostics. It consisted of a non contact IR based temperature control system for RT-PCR process and an optical detection system for on chip detection. It has been interfaced to perform both RT-PCR as well as chemiluminescence assays in sequence. It can detect HIV using p24 and gp120 markers [27]. A microfluidic single cell impedance cytometer was demonstrated to perform white blood cell differential count. The device is label free and could perform the analysis at high speed. It consists of micro electrodes which are used to measure the impedance of each single cell at two frequencies. The device demonstrated as a potential microcytometer for a POCT blood diagnostics [28].

Common Barrier in the technology

The biggest non technical hurdle to the implementation of POCT is the regulations laid down by the federal and the state government bodies such as FDA. The degree of oversight depends on the complexity of the test. For a low complexity test like pregnancy test, the patient can perform the test on their own specimens but for a complex test, there is a very high degree of regulatory requirements [7]. Another important problem is designing devices for a poor resource setting. A comparison of different design constrains for various resources is given in table 3 [29]. Table 3: Range of appropriate LOC procedures for different settings.
Storage and transportati on 4 to 25 C Sample pretreatme nt Off-chip technici an LOC procedures Fluid Fluid actuation control mixing Signal detectio n Expensi ve bulky, w/ ground electricit y Portable detector Bulky detector, w/ ground electricit y disposal

Settings

Material & manufacturi ng High cost & low cost

Highincome centralis ed Highincome Point-ofcare lowincome centralis ed

Electrokinet ic; pneumatic

Technicia n; machine; on-chip valves

Active or Passiv e

Incinerati on

Low cost

4 to 25 C Rough handling 4 to 40 C

On-chip

Pneumatic; capillary

Passive

Passiv e

Selfcontained

Low cost

Off-chip technici an

Electrokinet ic; pneumatic

Technicia n; machine; on-chip valves

Active or passiv e

Incinerati on

Another hurdle is the miniaturisation of the device and the technology which it should be easy to conduct, fast and should have high sensitivity and accurate result. Another hurdle is making the device and the technology extremely simple to use. This has proved to be the main downfall otherwise promising LOAC devices, it terms of their ability to be used as POCT devices [7].

Some possible solutions

New materials should be exploited for planar microfluidics such as paper as substrate. The material should be easy to integrate open channel system with capillaries and also should autonomously enable droplet based movement of fluids[7].New detection techniques should be developed which can be integrated on the chip or can be easily integrated such as OLED based detector [30].

Conclusion
POCT devices have the potential to solve the global health healthcare problem to a certain extent. But at present, there are only a few POCT devices which are cheap and can be afforded by the developing countries. The design constrains of the poor resource setting of developing countries add to the technical challenges. As a reason, technical innovations are required in the LOAC technology to solve these problems. One solution may be use of passive microchip such as lab on paper [31] and non- and minimally instrumented microfludic based diagnostic devices [32] and innovations in the detection technique which usually requires optical detector. Integrated biosensors can also be used as a detector. With the current trend of technical innovations and increasing number of industries taking interest in LOAC technology, in the next decade, the POCT devices may be able make a difference in the lives of the people around the world.

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