使用四种商用DNA提取试剂盒通过实时PCR检测农场环境样本中的肉毒梭菌III型。
Detection of Clostridium botulinum group III in environmental samples from farms by real-time PCR using four commercial DNA extraction kits.
作者信息
Le Maréchal Caroline, Fourour Sarah, Ballan Valentine, Rouxel Sandra, Souillard Rozenn, Chemaly Marianne
机构信息
ANSES, Laboratoire de Ploufragan-Plouzané, Unité Hygiène et Qualité des Produits Avicoles et Porcins, Université Bretagne-Loire, BP 53, 22440, Ploufragan, France.
ANSES, Laboratoire de Ploufragan-Plouzané, Unité d'Épidémiologie et Bien-être en Aviculture et Cuniculture, Université Bretagne-Loire, BP 53, 22440, Ploufragan, France.
出版信息
BMC Res Notes. 2018 Jul 4;11(1):441. doi: 10.1186/s13104-018-3549-5.
OBJECTIVES
Few studies have tested DNA extraction methods to optimize the detection of Clostridium botulinum in environmental samples that can be collected during animal botulism outbreaks. In this study, we evaluated four commercial DNA extraction kits for the detection of C. botulinum group III in 82 various environmental samples (9 manure, 53 swabs, 3 insects, 8 water, 1 silage and 8 soil samples) collected in a context of animal botulism outbreaks.
RESULTS
The PowerSoil kit was the most efficient for almost all matrices (83.6% of the 73 tested samples), except manure for which the NucleoSpin Soil kit was the most efficient. The NucleoSpin Soil kit enabled detection in 75.3%, the QIAamp DNA Mini Kit in 68.5%, and the QIAamp Fast DNA Stool Mini Kit in 45.2%. However, the NucleoSpin Soil kit detected C. botulinum in 9 of the 9 manure samples tested, while the PowerSoil kit found C. botulinum in only two samples, and the other two kits in none of the samples. This study showed that PowerSoil can be recommended for DNA extraction from environmental samples except for manure, for which the NucleoSpin Soil kit appeared to be far more appropriate.
目的
很少有研究测试过DNA提取方法，以优化在动物肉毒中毒暴发期间可采集的环境样本中肉毒梭菌的检测。在本研究中，我们评估了四种商用DNA提取试剂盒，用于检测在动物肉毒中毒暴发背景下采集的82份各种环境样本（9份粪便、53份拭子、3份昆虫、8份水、1份青贮饲料和8份土壤样本）中的III型肉毒梭菌。
结果
PowerSoil试剂盒对几乎所有基质（73份测试样本中的83.6%）最为有效，但粪便样本除外，对于粪便样本，NucleoSpin Soil试剂盒最为有效。NucleoSpin Soil试剂盒的检出率为75.3%，QIAamp DNA Mini试剂盒为68.5%，QIAamp Fast DNA Stool Mini试剂盒为45.2%。然而，NucleoSpin Soil试剂盒在9份测试粪便样本中的9份中检测到了肉毒梭菌，而PowerSoil试剂盒仅在两份样本中检测到肉毒梭菌，其他两种试剂盒在任何样本中均未检测到。本研究表明，除粪便外，PowerSoil可推荐用于从环境样本中提取DNA，对于粪便样本，NucleoSpin Soil试剂盒似乎更为合适。
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