全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

食品中克罗诺杆菌(原阪崎肠杆菌)双重PCR检测方法研究

DOI: 10.7685/j.issn.1000-2030.2012.01.020, PP. 113-118

Keywords: 克罗诺杆菌(原阪崎肠杆菌),双重PCR,食品检测

Full-Text   Cite this paper   Add to My Lib

Abstract:

根据克罗诺杆菌(Cronobacterspp.)16SrRNA基因以及局部大分子合成(MMS)操纵子特异序列设计2对引物,经反应体系和条件优化,建立了双重PCR检测方法。特异性检测结果显示,Cronobacterspp.菌株PCR扩增均可见2条特异性条带,而其他菌株PCR扩增均为阴性。纯菌检测双重PCR的灵敏度为6.3×103CFU?mL-1,而相对应单重PCR的灵敏度分别为6.3×101CFU?mL-1和6.3×103CFU?mL-1;人工污染的食品样品(奶粉、牛奶、鸡肉)在经过24h增菌后,检测限均可达100CFU?mL-1或100CFU?g-1。在鼠伤寒沙门氏菌(Salmonellatyphimurium)存在的条件下,双重PCR的检测限没有受到影响。表明本试验建立的双重PCR检测方法具有很好的特异性和灵敏度,能克服食品样品基质及杂菌的干扰,可应用于食品中Cronobacterspp.的检测。

References

[1]  Zhou Y,Wu Q,Xu X,et al.Development of an immobilization and detection method of Enterobacter sakazakii from powdered infant formula[J].Food Microbiology,2008,25:648-652
[2]  叶应旺,吴清平,郭伟鹏,等.种特异性PCR快速检测奶粉中阪崎肠杆菌研究[J].微生物学通报,2007,34(6):1192-1197
[3]  Iversen C,Forsythe S.Isolation of Enterobacter sakazakii and other Enterobacteriaceae from powdered infant formula milk and related products[J].Food Microbiology,2004,21:771-777
[4]  FAO/WHO.Enterobacter sakazakii and other microorganisms in powdered infant formula:meeting report.MRA Series 6 .(2004) .http://www.who.int/foodsafety/micro/jemra/meetings/feb2004/en/
[5]  Kong R Y C,Lee S K Y,Law T W F,et al.Rapid detected of six types of bacterial pathogens in marine waters by multiplex PCR[J].Water Research,2002,36(11):2802-2812
[6]  Healy B,Cooney S,O’Brien S,et al.Cronobacter(Enterobacter sakazakii):an opportunistic foodborne pathogen[J].Foodborne Pathog Dis,2010,7(4):339-350
[7]  FAO/WHO.Enterobacter sakazakii(Cronobacter spp.)in powdered follow-up formulae .Rome,Italy:Microbiological Risk Assessment Series No.15,2008:7-8
[8]  Friedemann M.Enterobacter sakazakii in food and beverages(other than infant formula and milk powder)[J].International Journal of Food Microbiology,2007,116(1):1-10
[9]  Kandhai M C,Reij M W,Gorris L G,et al.Occurrence of Enterobacter sakazakii in food production environments and households[J].The Lancet,2004,363(9402):39-40
[10]  Kandhai M C,Heuvelink A E,Reij M W,et al.A study into the occurrence of Cronobacter spp.in the Netherlands between 2001 and 2005[J].Food Control,2010,21(8):1127-1136
[11]  Malorny B,Wagener M.Detection of Enterobacter sakazakii strains by real-time PCR[J].Journal of Food Protection,2005,68(8):1623-1627
[12]  Lehner A,Tasara T,Stephan R. 16S rRNA gene based analysis of strains from different sources and development of a PCR assay for identification[J]. BMC Microbiology,2004,4(1):43
[13]  高旗利,张霞,罗茂凰,等.奶粉中阪崎肠杆菌PCR检测方法研究[J].检验检疫科学,2005,15(4):4-8
[14]  Nair M K M,Venkitanarayanan K S.Cloning and sequencing of the ompA gene of Enterobacter sakazakii and development of an ompA-targeted PCR for rapid detection of Enterobacter sakazakii in infant formula[J].Appl Environ Microbiol,2006,72(4):2539-2546
[15]  Lehner A,Grimm M,Rattei T,et al.Cloning and characterization of Enterobacter sakazakii pigment genes and in situ spectroscopic analysis of the pigment[J].FEMS Microbiology Letters,2006,265(2):244-246
[16]  叶应旺,吴清平,郭伟鹏,等.PCR快速检测奶粉中阪崎肠杆菌研究[J].中国卫生检验杂志,2007,17(3):422-424,467
[17]  Seo K H,Brackett R E.Rapid,specific detection of Enterobacter sakazakii in infant formula using a real-time PCR assay[J].Journal of Food Protection,2005,68(1):59-63
[18]  Ye Y,Wu Q,Yao L,et al.A comparison of polymerase chain reaction and international organization for standardization methods for determination of Enterobacter sakazakii contamination of infant formulas from Chinese mainland markets[J].Foodborne Pathog Dis,2009,6(10):1229-1234
[19]  Kang S E,Nam Y S,Hong K W.Rapid detection of Enterobacter sakazakii using TaqMan real-time PCR assay[J].J Microbiol Biotechnol,2007,17(3):516-519
[20]  侯俊如,张伟,张会彦,等.坂崎肠杆菌DNA提取方法比较和增菌研究[J].微生物学杂志,2007,27(4):97-100

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133