title: An improved RT-IPCR for detection of pyrene and related polycyclic aromatic hydrocarbons.
abstract: Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous homogeneous chemicals which are well known by carcinogens, mutagens and endocrine disorder. Here, an improved real-time immuno-PCR (RT-IPCR) was developed for detection of pyrene and its homologs in water samples. The PAHs in sample compete with pyrene-modified DNA to bind with monoclonal antibody (McAb) coated on PCR plate. The reporter DNA was exponentially amplified by real-time PCR instrument using Fast Start universal SYBR Green Master (ROX) kit. Only two reaction steps were needed to accomplish the detection. The assay had a good linear range from 5 pmol L(-1) to 5 nmol L(-1) with a detection limit of 3.5 pmol L(-1). For application assay, the average recoveries from tap water, lake water and mineral water were 98.4%, 98.2% and 99.7%, respectively which showed a good correlation (R(2)=0.9906) with those from GC-MS. The results indicated that the improved RT-IPCR seems to be a potential method for simple and ultrasensitive detection of pyrene and some homologues in environment water samples.
authors.affiliation: Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses/College of Veterinary Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130062, PR China; State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors.forename: X Y
authors.initials: XY
authors.lastname: Meng
authors.affiliation: Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses/College of Veterinary Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130062, PR China.
authors.forename: Y S
authors.initials: YS
authors.lastname: Li
authors.affiliation: Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses/College of Veterinary Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130062, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, People's Republic of China. Electronic address: zhouyurunye@sina.com.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses/College of Veterinary Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130062, PR China; Biological Engineering Department, Jilin Business and Technology College, Changchun 130507, PR China.
authors.forename: Y
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses/College of Veterinary Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130062, PR China.
authors.forename: B
authors.initials: B
authors.lastname: Qiao
authors.affiliation: Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses/College of Veterinary Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130062, PR China.
authors.forename: C C
authors.initials: CC
authors.lastname: Si
authors.affiliation: Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses/College of Veterinary Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130062, PR China.
authors.forename: P
authors.initials: P
authors.lastname: Hu
authors.affiliation: Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses/College of Veterinary Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130062, PR China.
authors.forename: S Y
authors.initials: SY
authors.lastname: Lu
authors.affiliation: Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses/College of Veterinary Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130062, PR China.
authors.forename: H L
authors.initials: HL
authors.lastname: Ren
authors.affiliation: Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses/College of Veterinary Medicine, The First Hospital of Jilin University, Jilin University, Changchun 130062, PR China.
authors.forename: Z S
authors.initials: ZS
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors.forename: H J
authors.initials: HJ
authors.lastname: Qiu
authors.affiliation: Production Quality Test Institute of Jilin Province, Changchun 130022, PR China.
authors.forename: J Q
authors.initials: JQ
authors.lastname: Liu
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D015374:Biosensing Techniques; D004247:DNA / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005618:Fresh Water / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry; D011084:Polycyclic Aromatic Hydrocarbons / Q000302:isolation & purification*; D011721:Pyrenes / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D014874:Water Pollutants, Chemical / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D011084:Polycyclic Aromatic Hydrocarbons; D011721:Pyrenes; D014874:Water Pollutants, Chemical; D004247:DNA; C030984:pyrene
keywords: Detection; Improved RT-IPCR; PAHs