Preparation and application of Vibrio parahaemolyticus using immunomagnetic beads

The objective of this study was to use Immunomagnetic beads (IMBs) to detect Vibrio parahamolyticus.IMBs were synthesized using anti-V.parahaemolyticus polyclonal antibody (commercial antibody R1 and self-regulating antibody R2) and magnetic beads coated with activated carboxyl groups and tosylated surfaces groups (two carboxyl IMBs IMB-C1 and IMB-C2;three toslated IMBs IMB-T1, IMB-T2 and IMB-T3 which coated by the secondary antibody and polyclonal antibody) .After the IMS process, one part of suspension was plated on TCBS to count the colony, the other to extract DNA with thermal cracking for Loop-mediated Isothermal Amplification (LAMP) .The antibody concentration per surface area (AC/SA) was used for evaluation the properties of IMBs.The result showed that the AC/SA of the carboxyl IMBs are higher than toslated IMBs (34.25fg/μm2 and 36.73fg/μm2) .And the carboxyl IMBs captured more V.parahaemolyticus during the IMS process than toslated IMBs, when the pure culture concentration was about 4.32logCFU/mL (p<0.05) .After the IMS process, approximately 79%and 94%of the cells in suspension were captured by the carboxyl beads with commercial and self-regulating antibodies.And, there were not obviously different about the capturing rate between the directed and indirected conjugation methods, compared the IMB-T3 and IMB-T2.The LAMP results showed that the molecule detection method was not influenced by the IMBs-cell complex.