title: Magnetic beads-based chemiluminescent assay for ultrasensitive detection of pseudorabies virus.
abstract: A rapid, ultrasensitive and economical Pseudorabies virus (PRV) detection system based on magnetic beads (MBs) and chemiluminescence was developed in this paper. The carboxyl functionalized MBs (MBs-COOH) were covalently coupled with aminated DNA probes for capturing PRV biotinylated amplicon, the product of polymerase chain reaction (PCR). Agarose gel electrophoresis analysis approved the reliability of biotinylated amplicon. The MBs composites were incubated with alkaline phosphatase labeled streptavidin (ALP-SA) and chemiluminescene was determined by subsequently adding 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy)phenyl-1,2-dioxetane (AMPPD). The optimal conditions of the PRV detection method were 10 microM for probe concentration, 50 degrees C for hybridization temperature and 30 min for hybridization time. The limit of detection (LOD) was as low as 100 amol/5 pM of amplicon which proved that this approach for PRV detection was ultrasensitive.
authors.forename: Haowen
authors.initials: H
authors.lastname: Yang
authors.forename: Yafen
authors.initials: Y
authors.lastname: Guo
authors.forename: Song
authors.initials: S
authors.lastname: Li
authors.forename: Ganqiu
authors.initials: G
authors.lastname: Lan
authors.forename: Qinyang
authors.initials: Q
authors.lastname: Jiang
authors.forename: Xiurong
authors.initials: X
authors.lastname: Yang
authors.forename: Jing
authors.initials: J
authors.lastname: Fan
authors.forename: Zeeshan
authors.initials: Z
authors.lastname: Ali
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Tang
authors.forename: Xianbo
authors.initials: X
authors.lastname: Mou
authors.forename: Hongna
authors.initials: H
authors.lastname: Liu
authors.forename: Muhammad Ali A
authors.initials: MA
authors.lastname: Shah
authors.forename: Si
authors.initials: S
authors.lastname: Jin
authors.forename: Hesheng
authors.initials: H
authors.lastname: Jiang
authors.forename: Zhiyang
authors.initials: Z
authors.lastname: Li
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D015342:DNA Probes; D004587:Electrophoresis, Agar Gel; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D057230:Limit of Detection; D049449:Luminescence; D008280:Magnetics; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D015342:DNA Probes