An improved RT-IPCR for detection of pyrene and related polycyclic aromatic hydrocarbons

• Published in: Biosensors & bioelectronics 2016-04, Vol.78, p.194-199
• Main Authors: Meng, X.Y., Li, Y.S., Zhou, Y., Sun, Y., Qiao, B., Si, C.C., Hu, P., Lu, S.Y., Ren, H.L., Liu, Z.S., Qiu, H.J., Liu, J.Q.
• Format: Article
• Language: English
• Subjects: Antibodies, Monoclonal - chemistry; Assaying; Biosensing Techniques; Biosensors; Carcinogens; Deoxyribonucleic acid; Detection; DNA - chemistry; Enzyme-Linked Immunosorbent Assay - methods; Fresh Water - chemistry; Gas Chromatography-Mass Spectrometry; Homology; Improved RT-IPCR; PAHs; Polycyclic aromatic hydrocarbons; Polycyclic Aromatic Hydrocarbons - isolation & purification; Pyrenes; Pyrenes - isolation & purification; Real time; Real-Time Polymerase Chain Reaction - methods; Water Pollutants, Chemical - isolation & purification
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2015.11.037

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 5pmolL−1 to 5nmolL−1 with a detection limit of 3.5pmolL−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 (R2=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. •An improved RT-IPCR was developed for ultrasensitive detection of PAHs.•The limit of detection could be as low as 3.5pmolL−1.•Only two reaction steps were needed to accomplish the assay.•The assay has advantage in simplicity, low cost and speed compared with traditional RT-IPCR.