Application on Rapid Detection of Salmonellla spp in food by pyrosequencing technology

Abstract

Abstract:

To develop a technique for detecting and identifying Salmonellla spp by using pyrosequencing technology. According to virulence-specific target gene sequence analysis, this study designed specific PCR primers and sequencing primers respectively for pyrosequencing, and pyrosequencing reaction conditions were optimized. The results show that this method is the traditional national standard method of enrichment as a precondition before, while eliminating the tedious process of post-biochemical validation, greatly reducing the detection time from 4 or 7 days to 16 hours and accuracy consistent with the traditional biochemical validation. The study established Pyrosequencing detection method is effective, fast, accurate and simple operation for fast food-borne pathogen detection and identification of precise opened up broad prospects.

CLC Number:

Q781

References

[1] 牛天贵,张宝芹.食品微生物检验[M].北京:中国计量出版社,2003.9

[2] Barbara M.L, Tony C.B, Grahame W.G. The Microbiological Safety and Quality of Food[M]. Gaithersburg, Maryland, USA: Aspen publishers, 2000:374-886.

[3] GB 4789.4-2010,食品安全国家标准食品微生物学检验沙门氏菌检验[S].

[4] 邝良德,朱晓霞等.沙门氏菌快速检测技术概述[J].湖北农业科学,2011,50(2):226-232.

[5] 孙圆圆,赵鹏等.沙门氏菌检测方法研究进展[J].疾病防治,2011,38(1):218-221.

[6] 张胜凯,张先舟.环介导等温扩增技术检测沙门氏菌反应体系的优化[J].安徽农业科学,2010,38(22):11727-11729.

[7] 黄嫦娇,黄晓蓉.全自动荧光酶联免疫方法检测食品中沙门氏茵[J].安徽农业科学,2010,38(10):5320-5321.

[8] 陈之遥,周国华.焦测序技术的研究进展[J].现代生物医学进展,2008,8(8):1573-1598.

[9] Nian-cai Peng,Chun-lin Wang.Asymmetric PCR method in generation of HBV ssDNA for pyrosequencing[J].

Academic Journal of Xi’an Jiaotong University,2009,21(1):54-56.

[10] Zhi-Jun Yang,Mei-Zeng Tu.Comparison of amplicon-sequencing, pyrosequencing and real-time PCR for detection of YMDD mutants in patients with chronic hepatitis B[J]. World Journal of Gastroenterology,2006,12(44):7192-7196.

[11] 徐保红,李秀娟.PCR-焦磷酸测序法快速鉴定副溶血性弧菌[J]. 预防医学情报杂志,2011,27(1):78-80.

[12] 公衍文,任绪义.焦磷酸测序法鉴定临床常见病原菌[J]. 重庆医学,2010,39(22):3009-3010.

[13] Baback Gharizadeh, Biying Zheng, Michael Akhras, el al. Sentinel-base DNA genotyping using multiple sequencing primers for high-risk human papillomaviruses[J]. Molecular and Cellular Probes, 2006, 23:1-9.

[14] Ruth Ann Luna, Lea R. Fasciano, el al. DNA Pyrosequencing-Based Bacterial Pathogen Identi?cation in a Pediatric Hospital Setting[J]. American Society for Microbiology,2007,45(9): 2985–2992.

[15] 程绍辉,梁明华等. 焦磷酸测序技术在确认北京严重急性呼吸综合征 ( SARS) 病毒株并检测基因突变中的应用[J].病毒学报,2005,21(3)168-172.