Real-time immuno-PCR for ultrasensitive detection of pyrene and other homologous PAHs

被引：26

作者：

Meng, X. Y. ^{[1
]}

Li, Y. S. ^{[1
]}

Zhou, Y. ^{[1
]}

Zhang, Y. Y. ^{[1
]}

Qiao, B. ^{[1
]}

Sun, Y. ^{[1
]}

Yang, L. ^{[1
]}

Hu, P. ^{[1
]}

Lu, S. Y. ^{[1
]}

Ren, H. L. ^{[1
]}

Zhang, J. H. ^{[1
]}

Wang, X. R. ^{[1
]}

Liu, Z. S. ^{[1
]}

机构：

[1] Jilin Univ, Coll Vet Med, Inst Zoonosis, Key Lab Zoonosis,Minist Educ, Changchun 130062, Peoples R China

来源：

BIOSENSORS & BIOELECTRONICS | 2015年 / 70卷

基金：

中国国家自然科学基金;

关键词：

PAHs; Reporter DNA; RT-IPCR; Water; POLYCYCLIC AROMATIC-HYDROCARBONS; TEST KITS; IDENTIFICATION; ASSAY;

D O I：

10.1016/j.bios.2015.03.019

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

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-1 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. (C) 2015 Elsevier B.V. All rights reserved.

引用

页码：42 / 47

页数：6

共 32 条

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