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Development of a multiplex PCR assay for rapid and simultaneous detection of four genera of fish pathogenic bacteria
Zhang, D. F.1,2; Zhang, Q. Q.2; Li, A. H.2
2014-11-01
Source PublicationLETTERS IN APPLIED MICROBIOLOGY
ISSN0266-8254
Volume59Issue:5Pages:471-478
AbstractSpecies of genus Aeromonas, Vibrio, Edwardsiella and Streptococcus are the most common fish pathogenic bacteria that cause economically devastating losses in aquaculture. A multiplex polymerase chain reaction (mPCR) was developed for the simultaneous detection and differentiation of the four genera of fish pathogenic bacteria. Through the use of genus-specific primers instead of species-specific ones, the current mPCR covered much more target bacterial species compared with previously reported species-specific mPCR methods. The specificity of the four putative genus-specific primers was validated experimentally while used exclusively (uniplex PCR) or combined (mPCR) against bacterial genomic DNA templates of the target bacteria and nontarget bacteria. The PCR amplicons for the following genera were obtained as expected: Aeromonas (875bp), Vibrio (524bp), Edwardsiella (302bp) and Streptococcus (197bp), and the fragments could be separated clearly on the agarose gel electrophoresis. The mPCR did not produce nonspecific amplification products when used to amplify 21 nontarget species of bacteria. The mPCR detection limits for each target bacterial genera were 50 colony-forming units (CFU) in pure culture and 100CFU in fish tissue samples. In conclusion, the mPCR assay was proven to be a powerful alternative to the conventional culture-based method, given its rapid, specific, sensitive and reliable detection of target pathogens.
SubtypeArticle
KeywordAquaculture Detection Fish Pathogenic Bacteria Genus-specific Primers Multiplex Pcr
DOI10.1111/lam.12303
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaBiotechnology & Applied Microbiology ; Microbiology
WOS SubjectBiotechnology & Applied Microbiology ; Microbiology
WOS IDWOS:000343871400003
WOS KeywordPOLYMERASE-CHAIN-REACTION ; STREPTOCOCCUS-AGALACTIAE ; EDWARDSIELLA-ICTALURI ; IN-VITRO ; AEROMONAS ; CHINA ; SEQUENCES ; STRAINS ; TILAPIA ; TARDA
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Cited Times:7[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/20329
Collection鱼类生物学及渔业生物技术研究中心_期刊论文
Affiliation1.Chinese Acad Fishery Sci, Pearl River Fisheries Res Inst, Guangzhou, Guangdong, Peoples R China
2.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
Zhang, D. F.,Zhang, Q. Q.,Li, A. H.. Development of a multiplex PCR assay for rapid and simultaneous detection of four genera of fish pathogenic bacteria[J]. LETTERS IN APPLIED MICROBIOLOGY,2014,59(5):471-478.
APA Zhang, D. F.,Zhang, Q. Q.,&Li, A. H..(2014).Development of a multiplex PCR assay for rapid and simultaneous detection of four genera of fish pathogenic bacteria.LETTERS IN APPLIED MICROBIOLOGY,59(5),471-478.
MLA Zhang, D. F.,et al."Development of a multiplex PCR assay for rapid and simultaneous detection of four genera of fish pathogenic bacteria".LETTERS IN APPLIED MICROBIOLOGY 59.5(2014):471-478.
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