Please use this identifier to cite or link to this item:
Appears in Collections:Aquaculture Journal Articles
Peer Review Status: Refereed
Title: Use of restriction enzyme fragment length polymorphism (RFLP) of the 16S-23S rRNA internal transcribed spacer region (ITS) for identification of fish mycobacteria
Authors: Pourahmad, Fazel
Richards, Randolph
Contact Email:
Keywords: 16S–23S rRNA internal transcribed spacer (ITS) region
Restriction enzyme fragment length polymorphism (RFLP)
Fish mycobacteria
Issue Date: Oct-2013
Publisher: Elsevier
Citation: Pourahmad F & Richards R (2013) Use of restriction enzyme fragment length polymorphism (RFLP) of the 16S-23S rRNA internal transcribed spacer region (ITS) for identification of fish mycobacteria, Aquaculture, 410-411, pp. 184-189.
Abstract: PCR targeting the 16S-23S rRNA gene internally transcribed spacer (ITS) region has been proposed as a rapid and reliable method for the detection of Mycobacterium species in human clinical specimens. Because of variation in ITS sequences amongst Mycobacterium species, a single PCR amplification can be used to differentiate slowly growing and rapidly growing species within this genus. In the present study, analysis by ITS-PCR and ITS-restriction fragment length polymorphism (RFLP) was found to be a useful, simple and rapid method compared to current molecular and phenotypic techniques. The ITS was amplified from 13 reference strains and 59 fish isolated mycobacteria using a set of published PCR primers. After PCR, the banding patterns generated allowed slowly growing mycobacteria to be differentiated from all other rapidly growing species, with the exception of Mycobacterium conceptionense. HaeIII was selected as one of two restriction enzymes that, together with the knowledge about amplicon sizes, would produce an acceptable level of discrimination in the resulting RLFP patterns, especially in the rapidly growing group of mycobacteria. After digestion with Sau96I, the amplified products of most isolates of Mycobacterium fortuitum, including subtypes II and V and those 2 isolates with new patterns (220, 100bp), presented identical or very similar patterns as obtained by HaeIII digestion. All isolates of Mycobacterium marinum, Mycobacterium chelonae and Mycobacterium gordonae, whose PCR products were not digested with HaeIII, produced two well-defined fragments with the Sau96I restriction enzyme.
Type: Journal Article
DOI Link:
Rights: The publisher does not allow this work to be made publicly available in this Repository. Please use the Request a Copy feature at the foot of the Repository record to request a copy directly from the author. You can only request a copy if you wish to use this work for your own research or private study.
Affiliation: Ilam University

Files in This Item:
File Description SizeFormat 
Aquaculture 2013.pdf461.38 kBAdobe PDFUnder Embargo until 31/12/2999     Request a copy

Note: If any of the files in this item are currently embargoed, you can request a copy directly from the author by clicking the padlock icon above. However, this facility is dependant on the depositor still being contactable at their original email address.

This item is protected by original copyright

Items in the Repository are protected by copyright, with all rights reserved, unless otherwise indicated.

If you believe that any material held in STORRE infringes copyright, please contact providing details and we will remove the Work from public display in STORRE and investigate your claim.