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.
Roh
Changhyun
C
Radiation Research Division for Biotechnology, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Sinjeong-dong, Jeongeup, Jeonbuk, South Korea. chroh@kaeri.re.kr
Lee
Ho-Young
HY
Kim
Sang-Eun
SE
Jo
Sung-Kee
SK
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