Abstract
Late blowing, a microbiological spoilage in hard and semi-hard cheese caused by Clostridium spores in raw milk, results in high economic losses for cheese producers. This study compared the sensitivity of the newly developed multiplex qPCR method which employing novel oligonucleotide primers and fluorescent TaqMan probes, and the culture-based most probable number (MPN) method in detecting the late blowing agent Clostridium species in traditional Turkish cheese. A total of 50 naturally contaminated cheese samples obtained from producers were analysed by both methods. Clostridium tyrobutyricum was the most common species occurring in 74% of the cheese samples, followed by C. butyricum and C. sporogenes occurring in 50% and 16% of the samples, respectively. The results of the two methods were consistent in 42 out of the 50 (84%) cheese samples. Our results indicate that the multiplex qPCR method is more sensitive than the MPN method. The multiplex qPCR method provided a favourable alternative to traditional cultural methods. This alternative molecular method has great potential in the laboratory and in the field for the rapid detection of late blowing of cheese samples.
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Data Availability
The datasets analysed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
We are grateful to the Ege University Planning and Monitoring Coordination of Organizational Development and Directorate of Library and Documentation for their support in editing and proofreading service of this study.
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This work was funded by the Scientific and Technological Research Council of Turkey (TUBITAK; Project No. 218Z113).
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All authors contributed to the study conception and design. All authors wrote the main manuscript text and prepared tables. All authors reviewed the manuscript.
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Aslı Şahiner declares that she has no conflict of interest. Sennur Çalışkan declares that she has no conflict of interest. Ece Halat declares that she has no conflict of interest.
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Şahiner, A., Çalışkan, S. & Halat, E. Quantitative Detection of Late Blowing Agents C. tyrobutyricum, C. butyricum, and C. sporogenes in Traditional Turkish Cheese by Multiplex Real-Time PCR. Food Anal. Methods 16, 781–786 (2023). https://doi.org/10.1007/s12161-023-02454-z
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DOI: https://doi.org/10.1007/s12161-023-02454-z