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Production of a Taq Man Real – Time PCR kit for rapid determination of microbial load in pasteurized milk

farhoudi, aida (2015) Production of a Taq Man Real – Time PCR kit for rapid determination of microbial load in pasteurized milk. Masters thesis, Gazvin University of Medical Science.

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Abstract

Milk is an excellent source of nutrient. it contains many valuable compounds that have an important role in human health. This vital food in suitable condition such as pH and Ingrediant like Protein and fat can be easily perishable. Consumption of milk with insufficient processing can cause food born illness. Therefore evaluation of hygienic control of milk is always important to make sure of its safety. Sevral Methods have used to detect and quantify pathogenic microorganisms. Conventional microbial count methods are based on Plate Counting Agar (PCA) in which bacteria can only be cultivated if their metabolic requirements is reproduced in vitro. So these methods can be time consuming and may takes 72 hours, and they are inappropriate in situations where rapid diagnostic decisions are required and several contiguous bacteria may lead to count a colony. Recently using molecular methods have been increased for rapid and accurate enumoration of bacteria. In this study, we designed primer set and a labeled prob with FAM and TAMRA for 16S rRNA region to set Taq Man Real Time PCR and optimized it to enumerate microbial load in pasteurized milk sample. And correlation of its result with cultural result was evaluated by Bland-Altman plot. According to the cultur result, the minimum and maximum was 0 and 10000, respectively.and for qPCR it was assessed 55 and 7071. Result indicated that, as an alternative method, Taq Man Real Time PCR provides a tool for rapid and accurate quantification of microbial load in pasteurized milk samples.and it is therefore suitable for high throughput testing.

Item Type: Thesis (Masters)
Subjects: Q Science > QR Microbiology
S Agriculture > S Agriculture (General)
Divisions: University Thesis > Faculty of Health > Health and Food Safety
Depositing User: Health School Students
Date Deposited: 20 Dec 2015 05:40
Last Modified: 07 Dec 2016 12:13
URI: http://eprints.qums.ac.ir/id/eprint/2915

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