title: Real-time immuno-PCR for ultrasensitive detection of pyrene and other homologous PAHs.
abstract: Polycyclic aromatic hydrocarbons (PAHs) are significant environmental pollutant that can lead to cancer and endocrine system disrupting. Here we developed a real-time immuno-PCR (RT-IPCR) assay based on a biotinylated reporter DNA system for ultrasensitive detection of pyrene (PYR) and homologous PAHs in water. The PAHs in sample compete with PYR-OVA coated on PCR plate to bind with monoclonal antibody (McAb). The biotinylated goat anti-mouse IgG (Bio-IgG) can be captured by the McAb bound with PYR-OVA. Then streptavidin is bound with biotin on Bio-IgG. Finally biotinylated reporter DNA is captured by the streptavidin and quantified by real-time PCR using FastStart universal SYBR Green Master (ROX) kit. The linear range of the assay was from 500 fmol L(-1) to 5 nmol L(-)) with a detection limit of 450 fmol L(-1). The average recoveries of PYR and homologous PAHs from lake water, tap water and commercial mineral water were 96.8%, 101.4% and 99.6% respectively, indicating that water samples had little interfere with the assay. The results demonstrated that the developed RT-IPCR might be a potential method for ultrasensitive detection of PYR and homologous PAHs in drinking and environment water sample.
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: X Y
authors: XY
authors: Meng
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: Y S
authors: YS
authors: Li
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China. Electronic address: zhouyu69@sina.com.
authors: Y
authors: Y
authors: Zhou
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: Y Y
authors: YY
authors: Zhang
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: B
authors: B
authors: Qiao
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: Y
authors: Y
authors: Sun
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: L
authors: L
authors: Yang
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: P
authors: P
authors: Hu
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: S Y
authors: SY
authors: Lu
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: H L
authors: HL
authors: Ren
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: J H
authors: JH
authors: Zhang
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: X R
authors: XR
authors: Wang
authors: Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors: Z S
authors: ZS
authors: Liu
mesh_terms: D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D004784:Environmental Monitoring / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008832:Microchemistry / Q000295:instrumentation; D011084:Polycyclic Aromatic Hydrocarbons / Q000032:analysis* / Q000276:immunology; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019809:Streptavidin / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D014874:Water Pollutants, Chemical / Q000032:analysis*
chemical_list: D011084:Polycyclic Aromatic Hydrocarbons; D014874:Water Pollutants, Chemical; D004247:DNA; D019809:Streptavidin
keywords: PAHs; RT-IPCR; Reporter DNA; Water