High-sensitivity capillary gel electrophoretic analysis of DNA fragments on an electrophoresis microchip using electrokinetic injection with transient isotachophoretic preconcentration

Xu, Zhongqi; Hirokawa, Takatsugu; Nishine, Tsutomu; Arai, Akihiro

doi:10.1016/s0021-9673(03)00053-0

Cited by 64 publications

(49 citation statements)

References 23 publications

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“…This is par- ticularly suited for preconcentration with indirect detection, whereby varying the separation electrolyte co-ion will also impact on detection performance. The same group has applied this method to the analysis of DNA and proteins in microchips using single-channel and crossgeometry microchips [69][70][71][72] with a 10-to 30-fold increase in sensitivity obtained.…”

Section: Isotachophoretic Stackingmentioning

confidence: 99%

Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips

2007

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Poor sensitivity is considered to be one of the major limitations of electrophoretic separation methods, particularly when compared to traditional liquid chromatographic techniques. To address this issue, various in-line preconcentration techniques have been developed over the past 15 years, ranging in power and complexity, and there are now a number of well understood approaches routinely capable of providing a 10,000- to 100,000-fold increase in sensitivity, as well as several that can be pushed above a million. Furthermore, these have been achieved with particularly troublesome and often difficult samples, such as those having high salinity from a biological or environmental origin. This review will discuss the most common methods for improving the sensitivity of CE, CEC and microchip version of these, with particular attention to those approaches developed over the last five years.

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Section: Isotachophoretic Stackingmentioning

confidence: 99%

Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips

2007

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“…The combination of these two fields requires researchers knowledgeable in both subjects, but many scientists lack such a comprehensive background. The relatively low sensitivity of CE analysis is the second limitation in the field of DNA analysis [136][137][138][139][140]. The double bond formed by the base group in DNA has a low UV-visible absorption, and a normal UV detector has a low sensitivity.…”

Section: Limitations Advantages and Future Development Of Ce Based Omentioning

confidence: 99%

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Lian

Zhao

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

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Rapid transmission, high morbidity, and mortality are the features of human infectious diseases caused by microorganisms, such as bacteria, fungi, and viruses. These diseases may lead within a short period of time to great personal and property losses, especially in regions where sanitation is poor. Thus, rapid diagnoses are vital for the prevention and therapeutic intervention of human infectious diseases. Several conventional methods are often used to diagnose infectious diseases, e.g. methods based on cultures or morphology, or biochemical tests based on metabonomics. Although traditional methods are considered gold standards and are used most frequently, they are laborious, time consuming, and tedious and cannot meet the demand for rapid diagnoses. Disease diagnosis using capillary electrophoresis methods has the advantages of high efficiency, high throughput, and high speed, and coupled with the different nucleic acid detection strategies overcomes the drawbacks of traditional identification methods, precluding many types of false positive and negative results. Therefore, this review focuses on the application of capillary electrophoresis based on nucleic detection to the diagnosis of human infectious diseases, and offers an introduction to the limitations, advantages, and future developments of this approach.

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“…Other applications were also reported [13][14][15][16]. In this study, the EKS technique on the single-channel microchip for DNA fragments [17] was firstly applied for identification of PCR samples. The technique was also achieved on the cross microchip in practice and computer-simulated.…”

Section: Introductionmentioning

confidence: 98%

“…On single-channel microchips the EKS mode could easily be adopted because the simplest chip arrangement is similar to that of a miniaturized capillary [17]. But the same preconcentration was not possible on a cross microchip since only the short sample zone at the cross part might be introduced into the separation channel by using the familiar injection and separation procedure.…”

Section: Introductionmentioning

confidence: 99%

Performance of electrokinetic supercharging for high‐sensitivity detection of DNA fragments in chip gel electrophoresis

Nishine

Arai

et al. 2004

Electrophoresis

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Chip gel electrophoresis was explored for high-sensitivity detection of DNA by combining electrokinetic injection with transient isotachophoresis preconcentration (here named electrokinetic supercharging (EKS)). Low concentrations (0.2 mg/L) of DNA sample could be detected without fluorescence labeling using a conventional UV detector (at 260 nm). On a single-channel microchip, identification of PCR product was performed by exploiting both external and internal calibration methods. The deviation between the two calibration methods was about 3.6%, and the identified DNA fragment size matched with the predicted size of the template DNA. On the cross microchip the EKS preconcentration has also been achieved when changing the injection reservoir differing from the one used previously. The procedure was computer-simulated and the influence of the voltage applied to two-side reservoirs on sample preconcentration and dilution was also discussed.

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High-sensitivity capillary gel electrophoretic analysis of DNA fragments on an electrophoresis microchip using electrokinetic injection with transient isotachophoretic preconcentration

Cited by 64 publications

References 23 publications

Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips

Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Performance of electrokinetic supercharging for high‐sensitivity detection of DNA fragments in chip gel electrophoresis

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