羽绒中亚硫酸盐还原梭状芽孢杆菌荧光PCR检测方法研究
2017-03-01何永盛林霖赖心田王坤吴佳辉陈国
何永盛+林霖+赖心田+王坤+吴佳辉+陈国培
摘要:现有标准对羽绒服装产品中亚硫酸盐还原梭状芽孢杆菌的检测存在检测周期长、结果缺乏确证手段等不足之处,需要建立更为快速准确的检测方法进行补充和完善。本研究建立了可用于亚硫酸盐还原梭状芽孢杆菌鉴定的实时荧光PCR检测方法,并对该方法的特异性和检出限进行了分析验证,结果显示建立的方法具有良好的特异性,检出限为20 CFU/mL。在50批次样品检测中,本研究建立的方法与国标检测方法具有较好的一致性,可以应用于羽绒服装样品的日常检测。
关键词:羽绒;亚硫酸盐还原梭状芽孢杆菌;实时荧光PCR
中图分类号:TS107 文献标志码:A
Research on Fluorescence PCR Method for Detecting Sulfite-reducing Clostridia in Down Products
Abstract: Sulfite-reducing clostridia is an important detection index for down products. Due to long testing period and insufficient accuracy, the existing standard test method should be further complemented and improved. Hence, it is necessary to develop a quick and accurate detection method. In this study, a fluorescence PCR method for detecting sulfite-reducing clostridia was established, and the sensitivity and detection limit of the method were analyzed and verified by subsequent experiments. The experiment results showed that the established method can distinguish sulfite-reducing clostridia from other bacteria effectively, and its detection limit reached 20 CFU/mL. The testing of fifty lots of down products verified that the established method is consistent with national standard test method and it can be applied to routine testing of down products.
Key words: down product; sulfite-reducing clostridia; fluorescence real-time PCR
我國是羽绒及其制品的生产、出口和消费大国,近年来羽绒的安全卫生问题越来越受到人们的关注。微生物卫生指标不合格是制约我国羽绒服装产品出口的一个重要因素。目前,我国和欧盟的相关标准都对羽绒产品的微生物限量提出了明确的要求。作为羽绒服装产品检测中的重要微生物指标之一,亚硫酸盐还原梭状芽孢杆菌对外界环境有极强的耐受能力,高温灭菌等一般的灭菌方法难以将其杀死,因此在检测当中往往成为导致羽绒服装产品不合格的主要污染菌。我国羽绒服装中针对该菌的检测多采用国家标准GB/T 14272 — 2011《羽绒服装》中规定的方法,主要通过测定平板上的黑点状菌落数目来判定该菌是否超标。这种检测方法虽然操作简单,但却也存在检测周期过长以及检测结果缺乏确证手段等不足之处,因此有必要建立更为快速准确的检测方法对其进行补充和完善。……
