title: [Detection of Prorocentrum minimum (Pavillard) Schiller with the electrochemiluminescence-molecular probe].
abstract: In this study, the electrochemiluminescence-molecular probe (ECL-MP) was established based on the sandwich hybridization integrated with nuclease protection assay (NPA-SH) improved by electrochemiluminescence (ECL). It can be used for detecting Prorocentrum minimum (Pavillard) Schiller qualitatively and quantitatively. NPA probes of P. minimum were designed based on the NPA-SH. After labeled with Ru (bpy)3(2+) and biotin, they bounded with streptavidin-coupled magnetic beads and generated electrochemiluminescence in an ECL analyzer. The ECL counts per second (CPS) was drawn against the cell number obtained from microscopy to establish a calibration curve of P. minimum. The results showed that the labeled NPA probes had good specificity and practicability, the optimal usage of magnetic beads was 4 microg for detecting 20 microL hybridization mixture, the detection range of P. minimum cell numbers was 6.25 x 10(2) - 4 x 10(4), and there was no significant difference between the data gained from ECL-MP and microscopy with 95% confidence level (t-test) when treated with cultured and mixed samples. ECL-MP was a convenient new tool for rapid assessment of P. minimum in marine environment.
authors.affiliation: College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China. myfavorite_1314@163.com
authors.forename: Xia
authors.initials: X
authors.lastname: Zhu
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhen
authors.forename: Tie-Zhu
authors.initials: TZ
authors.lastname: Mi
authors.forename: Zhi-Gang
authors.initials: ZG
authors.lastname: Yu
authors.forename: Zhen-Ming
authors.initials: ZM
authors.lastname: Chi
authors.forename: Xing-Lan
authors.initials: XL
authors.lastname: Lu
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation; D004141:Dinoflagellida / Q000302:isolation & purification*; D055664:Electrochemical Techniques; D057097:Harmful Algal Bloom; D008163:Luminescent Measurements / Q000295:instrumentation* / Q000379:methods*; D015336:Molecular Probe Techniques / Q000295:instrumentation*