Establishment of a Multiplex PCR Assay to Detect Five Major Freshwater Bacteria

Date
2017
Authors
Gao, Weihua
Ai, Kete
Luo, Kai
Huang, Tinghua
Yao, Min
Hu, Wei
Fang, Liu
Qi, Zhitao
Xu, Qiaoqing
Contributor
Advisor
Department
Instructor
Depositor
Speaker
Researcher
Consultant
Interviewer
Journal Title
Journal ISSN
Volume Title
Publisher
Volume
Number/Issue
Starting Page
Ending Page
Alternative Title
Abstract
A multiplex polymerase chain reaction (mPCR) method for simultaneous detection of Aeromonas hydrophila, Streptococcus agalactiae, Klebsiella pneumoniae, Edwardsiella tarda, and E. ictalur was developed to rapidly and accurately identify the five most common bacteria that infect aquatic animals. The expected amplicons for ahe2 gene of A. hydrophila, cpsE gene of S. agalactiae, khe gene of K. pneumoniae, mukF gene of E.tarda, and the serC gene of E. ictaluri were 853 bp, 685 bp, 428 bp, 356 bp, and 124 bp, respectively. In the single PCR assays, the minimum detectable DNA contents were 13.2 pg for A. hydrophila, 27.4 pg for S. agalactiae, 1.95 pg for K. pneumoniae, 1.63 pg for E. tarda, 1.02 pg for E. ictalur. The detection limits of the multiplex PCR were 0.66 ng, 1.91 ng, 0.68 ng, 0.41 ng, 0.71 ng for A. hydrophila, S. agalactiae, K. pneumoniae, E. tarda and E. ictalur, respectively. The established multiplex PCR is significant for the rapid detection of common pathogenic bacteria of aquatic animals and provides the basis for the diagnosis of fish diseases.
Description
Keywords
multiplex PCR, freshwater bacteria, detection
Citation
Extent
9 pages
Format
Geographic Location
Time Period
Related To
Rights
Rights Holder
Collections
Email libraryada-l@lists.hawaii.edu if you need this content in ADA-compliant format.