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用于检测食品、土壤和婴儿粪便中A、B和E型肉毒梭菌的聚合酶链反应。

Polymerase chain reaction for detection of Clostridium botulinum types A, B and E in food, soil and infant faeces.

作者信息

Szabo E A, Pemberton J M, Gibson A M, Eyles M J, Desmarchelier P M

机构信息

Department of Microbiology, University of Queensland, Australia.

出版信息

J Appl Bacteriol. 1994 Jun;76(6):539-45. doi: 10.1111/j.1365-2672.1994.tb01650.x.

Abstract

The application of the polymerase chain reaction (PCR) for detection of Clostridium botulinum types A, B and E in foods, environmental and clinical samples was evaluated and compared to the mouse bioassay. Samples inoculated with 10, 100 and 1000 spores of Cl. botulinum types A and B included pasteurized milk, UHT milk, infant formula, infant faeces, meat juice, canned tuna, mushrooms, blood sausage and soil. Clostridium botulinum type E spores were inoculated into fish eggs, canned tuna, picked herring, raw fish and soil at similar levels. Spores were added to 2.5 g of each sample with the exception of soil which was inoculated in 10 g samples. The presence of Cl. botulinum in sample enrichments was determined by both PCR and the bioassay. An overall correlation of 95.6% was observed between PCR results and the mouse bioassay. Of the total of 114 samples tested there was disparity between the mouse bioassay and the PCR in three samples of soil inoculated with 100 type A or E spores and 10 type B spores per 10 g, respectively, and two samples of infant faeces inoculated with 10 type A or B spores per 2.5 g. All of these samples gave negative animal results and positive PCR results.

摘要

评估了聚合酶链反应(PCR)在食品、环境和临床样本中检测A型、B型和E型肉毒梭菌的应用,并与小鼠生物测定法进行了比较。接种了10、100和1000个A型和B型肉毒梭菌孢子的样本包括巴氏杀菌牛奶、超高温瞬时灭菌牛奶、婴儿配方奶粉、婴儿粪便、肉汁、罐装金枪鱼、蘑菇、血肠和土壤。将E型肉毒梭菌孢子以相似水平接种到鱼卵、罐装金枪鱼、腌制鲱鱼、生鱼和土壤中。除土壤接种10 g样本外,将孢子添加到每个2.5 g的样本中。通过PCR和生物测定法确定样本富集物中肉毒梭菌的存在情况。观察到PCR结果与小鼠生物测定法之间的总体相关性为95.6%。在总共114个测试样本中,每10 g分别接种100个A型或E型孢子和10个B型孢子的三个土壤样本,以及每2.5 g接种10个A型或B型孢子的两个婴儿粪便样本,小鼠生物测定法和PCR结果存在差异。所有这些样本的动物检测结果均为阴性,而PCR结果为阳性。

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