Development of a Multiplex PCR to Identify Lactobacillus acidophilus NCFM, a Probiotic Organism

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dc.contributor.advisorDr. Lee-Ann Jaykus, Committee Memberen_US
dc.contributor.advisorDr. Sophia Kathariou, Committee Memberen_US
dc.contributor.advisorDr. Todd Klaenhammer, Committee Chairen_US
dc.contributor.authorSouther, Nicole Elaineen_US
dc.date.accessioned2010-04-02T17:54:15Z
dc.date.available2010-04-02T17:54:15Z
dc.date.issued2010-04-02
dc.degree.disciplineFood Scienceen_US
dc.degree.levelthesisen_US
dc.degree.nameMSen_US
dc.description.abstractLactobacillus acidophilus NCFM is a probiotic organism used in food products and dietary supplements since 1972. A recent study recovered strains of L. acidophilus from various food- and probiotic products currently being used in the food industry demonstrating a genetic fingerprint comparable to that of L. acidophilus NCFM. Reliable strain identification in commercial products is of paramount in regards to food safety and consistent probiotic features. A multiplex PCR system to identify Lb. acidophilus NCFM was established and used to distinguish this strain from other Lactobacillus species and Lactobacillus acidophilus strains. The genome of this organism revealed the presence of seven distinct transposase elements. Three of these genes were found in multiple, highly conserved copies ranging from two to six replicas, indicating relatively recent multiplication events. A total of four different primer sets were designed around four selected transposase genes to generate semi-conserved PCR products. Various strains of L. acidophilus, including NCFM, were tested with the primer sets. The multiplex reaction identified six of these strains as NCFM, or closely related to Lb. acidophilus NCFM. Five of the L. acidophilus strains showed distinguishing banding patterns. We were able to demonstrate the use of multiplex PCR to identify L. acidophilus NCFM and NCFM-related strains, and distinguish these from other species and closely-related strains of L. acidophilus.en_US
dc.identifier.otheretd-11212004-170111en_US
dc.identifier.urihttp://www.lib.ncsu.edu/resolver/1840.16/273
dc.rightsI hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to NC State University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report.en_US
dc.subjectmultiplex PCRen_US
dc.subjectstrain identificatitonen_US
dc.subjectprobiotic bacteriaen_US
dc.subjectbacteria identification techniqueen_US
dc.titleDevelopment of a Multiplex PCR to Identify Lactobacillus acidophilus NCFM, a Probiotic Organismen_US

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