Tecnología de los Alimentos, Facultad de Veterinaria, Universidad de Zaragoza, Zaragoza, Spain.
J Appl Microbiol. 2010 Feb;108(2):488-98. doi: 10.1111/j.1365-2672.2009.04435.x. Epub 2009 Jun 25.
The present work investigates the feasibility of using flow cytometry (FCM) combined with fluorescent-labelled specific polyclonal antibodies for the detection and presumptive identification of Clostridium tyrobutyricum spores in bovine milk.
Two fluorescent molecules (fluorescein isothiocyanate and Alexa Fluor 488) were conjugated to antispores polyclonal antibodies. Side scatter and forward scatter profiles of the Cl. tyrobutyricum spores marked with fluorescent antibodies permitted the detection of spores and differentiated them from other related microbial species. The detection limit of this method was 10(3) spores per 100 ml of milk, and results could be achieved in 2 h.
FCM combined with fluorochrome-conjugated antibodies, especially Alexa Fluor, could be an efficacious means to detect and provide presumptive identification of Cl. tyrobutyricum spores, as well as differentiation from other Clostridium species that can also cause late blowing in cheese.
This study describes the basis for the development of a method suitable for analysis of milk destined for cheese manufacture that would permit the detection of Cl. tyrobutyricum spores in a short period. This would enable the industry to use contaminated milk for dairy products other than cheese where Cl. tyrobutyricum does not cause a problem.
本研究旨在探讨使用流式细胞术（FCM）结合荧光标记的特异性多克隆抗体来检测和初步鉴定牛牛奶中酪丁酸梭菌孢子的可行性。
两种荧光分子（异硫氰酸荧光素和 Alexa Fluor 488）被连接到抗孢子多克隆抗体上。用荧光抗体标记的酪丁酸梭菌孢子的侧向散射和前向散射图谱允许检测孢子，并将其与其他相关微生物物种区分开来。该方法的检测限为每 100 毫升牛奶中 10³个孢子，结果可在 2 小时内获得。
FCM 结合荧光标记抗体，特别是 Alexa Fluor，可以有效地检测和初步鉴定酪丁酸梭菌孢子，并区分可能导致奶酪后期产气的其他梭菌属物种。
本研究为开发一种适用于分析用于奶酪生产的牛奶的方法奠定了基础，该方法可在短时间内检测到酪丁酸梭菌孢子。这将使该行业能够将受污染的牛奶用于不会引起酪丁酸梭菌问题的乳制品。
