title: Multiple Cross Displacement Amplification Combined with Gold Nanoparticle-Based Lateral Flow Biosensor for Detection ofVibrio parahaemolyticus.
abstract: Vibrio parahaemolyticus(V. parahaemolyticus) is a marine seafood-borne pathogen causing severe illnesses in humans and aquatic animals. In the present study, multiple cross displacement amplification was combined with a lateral flow biosensor (MCDA-LFB) to detect thetoxRgene ofV. parahaemolyticusin DNA extracts from pure cultures and spiked oyster homogenates. Amplification was carried out at a constant temperature (62°C) for only 30 min, and amplification products were directly applied to the biosensor. The entire process, including oyster homogenate processing (30 min), isothermal amplification (30 min) and results indicating (∼2 min), could be completed within 65 min. Amplification product was detectable from as little as 10 fg of pureV. parahaemolyticusDNA and from approximately 4.2 × 102CFU in 1 mL of oyster homogenate. No cross-reaction with otherVibriospecies and with non-Vibriospecies was observed. Therefore, the MCDA-LFB method established in the current report is suitable for the rapid screening ofV. parahaemolyticusin clinical, food, and environmental samples.
authors.affiliation: State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Chinese Center for Disease Control and Prevention Beijing, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Chinese Center for Disease Control and PreventionBeijing, China; Department of Microbiology, Guizhou Medical UniversityGuiyang, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: Changping District Center for Disease Control and Prevention Beijing, China.
authors.forename: Dongxun
authors.initials: D
authors.lastname: Li
authors.affiliation: Institute of Microbiology, Jilin Provincial Center for Disease Control and Prevention Changchun, China.
authors.forename: Kewei
authors.initials: K
authors.lastname: Li
authors.affiliation: State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Chinese Center for Disease Control and Prevention Beijing, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Chinese Center for Disease Control and Prevention Beijing, China.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Chinese Center for Disease Control and Prevention Beijing, China.
authors.forename: Changyun
authors.initials: C
authors.lastname: Ye
keywords: MCDA-LFB; Vibro parahaemolyticus; lateral flow biosensor; limit of detection; multiple cross displacement amplification