食品科学2017，Vol.38Issue(24)：260-264,5.DOI:10.7506/spkx1002-6630-201724042
采样方法对冷鲜鸡表面细菌DNA提取及高通量测序结果的影响
Effects of Different Sampling Methods on Microbial DNA Extraction from Chilled Chicken and the High-Throughput Sequencing of Amplification Products
摘要
Abstract
Objective: To compare the swabbing and rinsing methods used for sampling chilled chicken by evaluating their effects on microbial DNA extraction and subsequent high-throughput sequencing. Methods: Four chilled chickens were divided into equal halves and sampled by superficial swabbing and rinsing methods for each half, respectively. Microbial genomic DNA was extracted from the collected samples and the V3-V4 region of the bacterial 16S rRNA gene was amplified by PCR. The PCR products were then subjected to high-throughput sequencing on an Illumina HiSeq sequencing platform. The obtained sequences were processed and analyzed using QIIME and other softwares. Results: There was no difference in bacterial genomic DNA extraction or bacterial community richness, diversity and structures between the samples collected by the two methods (P > 0.05). High-throughput sequencing showed that the bacterial community in chilled chicken samples consisted mainly of 7 phyla and 10 genera. Proteobacteria was the dominant phylum, accounting for more than 70% of the bacterial community;Shewanella,Pseudomonas,Acinetobacter,Psychrobacter,and Brochothrix were the dominant genera, each representing 10%–20% of the bacterial community. Conclusions: The two sampling methods exhibit no obvious difference in bacterial DNA extraction from chilled chicken or high-throughput sequencing. However, compared with swabbing, rinsing has the advantages of easy operation and non-secondary contamination, so it is more practical in microbiological analysis of chilled chicken. The experiment also provides useful data for the study of bacteria on chilled chicken.关键词
冷鲜鸡/菌群结构/涂抹法/冲洗法/高通量测序Key words
chilled chicken/bacterial community/swabbing method/rinsing method/high-throughput sequencing分类
轻工纺织引用本文复制引用
肖英平,何祥祥,戴宝玲,桂国弘,唐标,杨华..采样方法对冷鲜鸡表面细菌DNA提取及高通量测序结果的影响[J].食品科学,2017,38(24):260-264,5.基金项目
浙江省公益技术运用研究项目(2016C32073) （2016C32073）
浙江省植物有害生物防控重点实验室——省部共建国家重点实验室培育基地项目(2010DS700124-ZM1608) （2010DS700124-ZM1608）
