A highly sensitive and selective viral protein detection method based on RNA oligonucleotide nanoparticle.
Globally, approximately 170 million people (representing approximately 3% of the population worldwide), are infected with hepatitis C virus (HCV) and at risk of serious liver disease, including chronic hepatitis. We propose a new quantum dots (QDs)-supported RNA oligonucleotide approach for the specific and sensitive detection of viral protein using a biochip. This method was developed by immobilizing a HCV nonstructural protein 5B (NS5B) on the surface of a glass chip via the formation of a covalent bond between an amine protein group and a ProLinker glass chip. The QDs-supported RNA oligonucleotide was conjugated via an amide formation reaction from coupling of a 5'-end-amine-modified RNA oligonucleotide on the surface of QDs displaying carboxyl groups via standard EDC coupling. The QDs-conjugated RNA oligonucleotide was interacted to immobilized viral protein NS5B on the biochip. The detection is based on the variation of signal of QDs-supported RNA oligonucleotide bound on an immobilized biochip. It was demonstrated that the value of the signal has a linear relationship with concentrations of the HCV NS5B viral protein in the 1 microg mL(-1) to 1 ng mL(-1) range with a detection limit of 1 ng mL(-1). The major advantages of this RNA-oligonucleotide nanoparticle assay are its good specificity, ease of performance, and ability to perform one-spot monitoring. The proposed method could be used as a general method of HCV detection and is expected to be applicable to other types of diseases as well.
Radiation Research Division for Biotechnology, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Sinjeong-dong, Jeongeup, Jeonbuk, South Korea. chroh@kaeri.re.kr
Changhyun
C
Roh
Ho-Young
HY
Lee
Sang-Eun
SE
Kim
Sung-Kee
SK
Jo
D015374:Biosensing Techniques / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D053758:Nanoparticles / Q000737:chemistry; D040081:Protein Array Analysis / Q000295:instrumentation*; D045663:Quantum Dots; D012313:RNA / Q000737:chemistry* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017361:Viral Nonstructural Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase
D012313:RNA; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase