三重PCR快速检测婴幼儿奶粉中的病原菌
|
摘要
食源性疾病是食品安全的主要问题,时刻威胁着人们的健康和生命。在食源性疾病中,细菌性食物中毒居首位。近年来,阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌等病原菌引起的奶粉中毒事件时有发生。因此,食源性病原菌的检测是食品卫生安全检测中一个重要的部分。
目前,对食品中病原菌的检测方法主要有传统的分离鉴定方法、免疫学方法和分子生物学方法。其中,传统检测方法操作繁琐,检测时间较长,灵敏度较低;免疫学方法的特异性和灵敏度不是很理想;分子生物学方法特异性较强、检测时间较短、灵敏度较高。
三重PCR是一种在同一反应体系中加入三对特异性引物,同时扩增出三条目的DNA片段的扩增方法。本文根据阪崎肠杆菌的外膜蛋白基因ompA、金黄色葡萄球菌的耐热核酸酶基因nuc、蜡样芽胞杆菌的溶血素基因hblA设计了3对特异性引物,进行三重PCR反应,可实现对阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌三种食源性病原菌的同时检测。对15株菌进行检测,结果4株阪崎肠杆菌、2株金黄色葡萄球菌、1株蜡样芽胞杆菌均扩增出了特异性的目的条带,而另外的8株其它病原菌均无扩增条带。将阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌的扩增产物测序序列在GeneBank上进行BLAST比对,同源性均达到99%以上。试验过程中通过对退火温度、3对引物、Mg~(2+)、dNTPs、EasyTaq DNA Polymerase等参数的优化,确定了三重PCR的反应体系和反应程序,其反应体系为:10×Easy Taq Buffer(-Mg~(2+))2.5μL,2.5 mM dNTPs 2.5μL,50 mM MgSO4 1.5μL,10μM ompA上下游引物各0.6μL、10μM nuc上下游引物各1.0μL、10μM hblA上下游引物各1.0μL,模板DNA各2μL,5 U/μL EasyTaq DNA Polymerase 0.3μL,ddH2O补足25μL;反应程序为:95℃预变性5 min;按照94℃45 s,61℃45 s,72℃45 s进行30个循环;最后72℃延伸10 min。
将阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌三种病原菌随机组合,均能扩增出特异性的目的条带。在最佳试验条件下,该三重PCR方法同时检测阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌三种病原菌纯培养的灵敏度是103 CFU/mL。采用无水乙醇、氨水、石油醚与试剂盒相结合的方法提取DNA,该三重PCR方法同时检测阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌三种病原菌在婴幼儿奶粉中的检出限是104 CFU/g。对实际样品进行检测,并将三重PCR方法与国标方法进行比较,三重PCR方法对阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌三种病原菌的敏感性均为100%;特异性分别为96.4%、96.6%、100%;符合率分别为96.7%、96.7%、100%。
本研究以婴幼儿奶粉中出现过的阪崎肠杆菌、金黄色葡萄球菌和蜡样芽胞杆菌为对象,建立了检测这三种病原菌的三重PCR方法,为同时检测食品中的阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌提供了新方法。
Food-borne diseases constantly threatening people’s health and lives is a major public health problem in food security. Especially, bacterial food poisoning is the main factor which causes the food poisoning. At present, the contaminated infant milk powder scandal owing to Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus happens occasionally. So the detection of food-borne pathogenic bacteria is a great of part in food and health security.
At present, the detection of food-borne pathogenic bacteria mainly uses the traditional isolation and biochemical identification method, the immunological methods and the methods in molecular biology. It is complicated operation, time consuming and low-sensitive for the traditional isolation and biochemical identification method. The specificity and sensibility of immunological methods are unsatisfactory. The methods in molecular biology are higher-specific, shorter-time and higher-sensitive.
Triplex PCR is an amplification method that can amplify three target sequences through adding to three pairs of specific primers. In this study, three pairs of specific primers were designed according to Enterobacter sakazakii ompA gene, Staphylococcus aureus nuc gene and Bacillus cereus hblA gene and triplex PCR was established to simultaneous diagnose three kinds of food-borne pathogens. The detecting results of 15 strains of bacteria are that 4 strains of Enterobacter sakazakii, 2 strains of Staphylococcus aureus and 1 strain of Bacillus cereus had amplified specific target channels, and another 8 strains of bacteria could not have amplified channels. The sequencing results of Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus were compared with the target sequences at GenBank and homologies of three pathogenic bacteria were all above 99%. The reaction system and reaction procedure of triplex PCR were determined through optimizing annealing temperature, three pairs of primers, Mg~(2+), dNTPs and EasyTaq DNA Polymerase. The reaction system was 25μL: 10×Easy Taq Buffer(-Mg~(2+)) 2.5μL, 2.5 mM dNTPs 2.5μL, 50 mM MgSO4 1.5μL, each 10μM ompA primer 0.6μL, each 10μM nuc primer 1.0μL, each 10μM hblA primer 1.0μL, each template DNA 2μL, 5 U/μL EasyTaq DNA Polymerase 0.3μL, ddH20 7μL. The reaction procedure was pre-degenerated at 95℃for 5 min, degenerated at 94℃for 45 s, annealing at 61℃for 45 s, extention at 72℃for 45 s, run 30 cycles, final extention at 72℃for 10 min.
Specific target channels could be amplified with random combinations of Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus. Under the above optimum experiment condition, the sensitivities of the triplex PCR for Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus were 103 CFU/mL. Extracting genomic DNA with absolute ethyl alcohol, ammonia water, petroleum ether and kit extraction, the detection limits of the triplex PCR for Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus in infant milk powder were 104 CFU/g. Compared with the national standard method, the sensitivities of triplex PCR to detect the actual samples for the three pathogenic bacteria were all 100%; the specificities of triplex PCR to detect the actual samples for the three pathogenic bacteria were 96.4%, 96.6%, 100%, respectively; the coincidence rate of triplex PCR to detect the actual samples for the three pathogenic bacteria were 96.7%, 96.7%, 100%, respectively.
In the study, triplex PCR assay had been established according to the food-borne pathogenic bacteria appeared in infant milk powder and it would provide a new method to simultaneously detect Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus.
目前,对食品中病原菌的检测方法主要有传统的分离鉴定方法、免疫学方法和分子生物学方法。其中,传统检测方法操作繁琐,检测时间较长,灵敏度较低;免疫学方法的特异性和灵敏度不是很理想;分子生物学方法特异性较强、检测时间较短、灵敏度较高。
三重PCR是一种在同一反应体系中加入三对特异性引物,同时扩增出三条目的DNA片段的扩增方法。本文根据阪崎肠杆菌的外膜蛋白基因ompA、金黄色葡萄球菌的耐热核酸酶基因nuc、蜡样芽胞杆菌的溶血素基因hblA设计了3对特异性引物,进行三重PCR反应,可实现对阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌三种食源性病原菌的同时检测。对15株菌进行检测,结果4株阪崎肠杆菌、2株金黄色葡萄球菌、1株蜡样芽胞杆菌均扩增出了特异性的目的条带,而另外的8株其它病原菌均无扩增条带。将阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌的扩增产物测序序列在GeneBank上进行BLAST比对,同源性均达到99%以上。试验过程中通过对退火温度、3对引物、Mg~(2+)、dNTPs、EasyTaq DNA Polymerase等参数的优化,确定了三重PCR的反应体系和反应程序,其反应体系为:10×Easy Taq Buffer(-Mg~(2+))2.5μL,2.5 mM dNTPs 2.5μL,50 mM MgSO4 1.5μL,10μM ompA上下游引物各0.6μL、10μM nuc上下游引物各1.0μL、10μM hblA上下游引物各1.0μL,模板DNA各2μL,5 U/μL EasyTaq DNA Polymerase 0.3μL,ddH2O补足25μL;反应程序为:95℃预变性5 min;按照94℃45 s,61℃45 s,72℃45 s进行30个循环;最后72℃延伸10 min。
将阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌三种病原菌随机组合,均能扩增出特异性的目的条带。在最佳试验条件下,该三重PCR方法同时检测阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌三种病原菌纯培养的灵敏度是103 CFU/mL。采用无水乙醇、氨水、石油醚与试剂盒相结合的方法提取DNA,该三重PCR方法同时检测阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌三种病原菌在婴幼儿奶粉中的检出限是104 CFU/g。对实际样品进行检测,并将三重PCR方法与国标方法进行比较,三重PCR方法对阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌三种病原菌的敏感性均为100%;特异性分别为96.4%、96.6%、100%;符合率分别为96.7%、96.7%、100%。
本研究以婴幼儿奶粉中出现过的阪崎肠杆菌、金黄色葡萄球菌和蜡样芽胞杆菌为对象,建立了检测这三种病原菌的三重PCR方法,为同时检测食品中的阪崎肠杆菌、金黄色葡萄球菌、蜡样芽胞杆菌提供了新方法。
Food-borne diseases constantly threatening people’s health and lives is a major public health problem in food security. Especially, bacterial food poisoning is the main factor which causes the food poisoning. At present, the contaminated infant milk powder scandal owing to Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus happens occasionally. So the detection of food-borne pathogenic bacteria is a great of part in food and health security.
At present, the detection of food-borne pathogenic bacteria mainly uses the traditional isolation and biochemical identification method, the immunological methods and the methods in molecular biology. It is complicated operation, time consuming and low-sensitive for the traditional isolation and biochemical identification method. The specificity and sensibility of immunological methods are unsatisfactory. The methods in molecular biology are higher-specific, shorter-time and higher-sensitive.
Triplex PCR is an amplification method that can amplify three target sequences through adding to three pairs of specific primers. In this study, three pairs of specific primers were designed according to Enterobacter sakazakii ompA gene, Staphylococcus aureus nuc gene and Bacillus cereus hblA gene and triplex PCR was established to simultaneous diagnose three kinds of food-borne pathogens. The detecting results of 15 strains of bacteria are that 4 strains of Enterobacter sakazakii, 2 strains of Staphylococcus aureus and 1 strain of Bacillus cereus had amplified specific target channels, and another 8 strains of bacteria could not have amplified channels. The sequencing results of Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus were compared with the target sequences at GenBank and homologies of three pathogenic bacteria were all above 99%. The reaction system and reaction procedure of triplex PCR were determined through optimizing annealing temperature, three pairs of primers, Mg~(2+), dNTPs and EasyTaq DNA Polymerase. The reaction system was 25μL: 10×Easy Taq Buffer(-Mg~(2+)) 2.5μL, 2.5 mM dNTPs 2.5μL, 50 mM MgSO4 1.5μL, each 10μM ompA primer 0.6μL, each 10μM nuc primer 1.0μL, each 10μM hblA primer 1.0μL, each template DNA 2μL, 5 U/μL EasyTaq DNA Polymerase 0.3μL, ddH20 7μL. The reaction procedure was pre-degenerated at 95℃for 5 min, degenerated at 94℃for 45 s, annealing at 61℃for 45 s, extention at 72℃for 45 s, run 30 cycles, final extention at 72℃for 10 min.
Specific target channels could be amplified with random combinations of Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus. Under the above optimum experiment condition, the sensitivities of the triplex PCR for Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus were 103 CFU/mL. Extracting genomic DNA with absolute ethyl alcohol, ammonia water, petroleum ether and kit extraction, the detection limits of the triplex PCR for Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus in infant milk powder were 104 CFU/g. Compared with the national standard method, the sensitivities of triplex PCR to detect the actual samples for the three pathogenic bacteria were all 100%; the specificities of triplex PCR to detect the actual samples for the three pathogenic bacteria were 96.4%, 96.6%, 100%, respectively; the coincidence rate of triplex PCR to detect the actual samples for the three pathogenic bacteria were 96.7%, 96.7%, 100%, respectively.
In the study, triplex PCR assay had been established according to the food-borne pathogenic bacteria appeared in infant milk powder and it would provide a new method to simultaneously detect Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus.
引文
[1]于艳霞.浅议食品安全和食源性疾病控制[J].中国医药指南. 2009(12): 185-186.
[2]刘秀梅.食源性疾病仍是中国头号食品安全问题[R].北京:中国疾病预防控制中心, 2006.
[3]汪清,葛秀清,黄祖华.细菌性食物中毒的现状及检验[J].中国动物检疫. 2004(4): 20-22.
[4]陈建琳,刘明辉.细菌性食物中毒流行趋势及预防对策[J].中国卫生检验杂志. 2002(4): 481.
[5]邱文毅,钱进,胡月珍,等.食源性疾病现状及国际航行船舶食品卫生监管建议[J].口岸卫生控制. 2011(1): 43-46.
[6]乔铁军,张国祥,王幼雄.食源性疾病及预防探讨[J].浙江预防医学. 2007(11): 35-39.
[7] Mead P S, Slutsker L, Dietz V, et al. Food-related illness and death in the United States[J]. Emerg Infect Dis. 1999, 5(5): 607-625.
[8] Schraff RL. Health-related costs from foodborne illness in the United States.Produce Safety Project [R]. Georgetown University, Washington. D.C. 2010.
[9] Mccabe-Sellers B J, Beattie S E. Food safety: emerging trends in foodborne illness surveillance and prevention[J]. J Am Diet Assoc. 2004, 104(11): 1708-1717.
[10]李建科,陈锦屏.我国食品安全现状与入世后的形势与对策[J].食品科学. 2003(8): 272-276.
[11]王艳君.冷却肉中致病菌的多重PCR检测技术研究[D].河南:河南农业大学, 2007.
[12]吴雅儿.因喂服染有蜡样芽胞杆菌奶粉引起婴儿腹泻1例[J].海峡预防医学杂志. 2003(5): 4.
[13]萨日娜.一起由金黄色葡萄球菌引起的奶粉食物中毒[J].中国卫生检验杂志. 2005(5): 636.
[14]叶红萍.从雅士利婴儿配方奶粉中检出蜡样芽胞杆菌[J].海峡预防医学杂志. 2008(1): 4.
[15] Lehner A, Stephan R. Microbiological, epidemiological, and food safety aspects of Enterobacter sakazakii[J]. J Food Prot. 2004, 67(12): 2850-2857.
[16] Nazarowec-White M, Farber J M. Enterobacter sakazakii: a review[J]. Int J Food Microbiol. 1997, 34(2): 103-113.
[17] Breeuwer P, Lardeau A, Peterz M, et al. Desiccation and heat tolerance of Enterobacter sakazakii[J]. J Appl Microbiol. 2003, 95(5): 967-973.
[18] Iversen C, Druggan P, Forsythe S. A selective differential medium for Enterobacter sakazakii, a preliminary study[J]. Int J Food Microbiol. 2004, 96(2): 133-139.
[19]张爱霞,生庆海,张涛.食品中阪崎肠杆菌分析及其检测[J].中国乳品工业. 2005(10): 43-45.
[20] Muytjens H L, Zanen H C, Sonderkamp H J, et al. Analysis of eight cases of neonatal meningitis and sepsis due to Enterobacter sakazakii[J]. J Clin Microbiol. 1983, 18(1): 115-120.
[21]徐英春,张小江,陈民钧,等.肠杆菌属的耐药调查及抗感染用药探讨[J].中华医院感染学杂志. 2001(3): 77-79.
[22] Enterobacter sakazakii infections associated with the use of powdered infant formula--Tennessee, 2001[J]. MMWR Morb Mortal Wkly Rep. 2002, 51(14): 297-300.
[23] Bowen A B, Braden C R. Invasive Enterobacter sakazakii disease in infants[J]. Emerg Infect Dis. 2006, 12(8): 1185-1189.
[24] Teramoto S, Tanabe Y, Okano E, et al. A first fatal neonatal case of Enterobacter sakazakii infection in Japan[J]. Pediatr Int. 2010, 52(2): 312-313.
[25] Pagotto F J, Nazarowec-White M, Bidawid S, et al. Enterobacter sakazakii: infectivity and enterotoxin production in vitro and in vivo[J]. J Food Prot. 2003, 66(3): 370-375.
[26] Townsend S, Caubilla B J, Loc-Carrillo C, et al. The presence of endotoxin in powdered infant formula milk and the influence of endotoxin and Enterobacter sakazakii on bacterial translocation in the infant rat[J]. Food Microbiol. 2007, 24(1): 67-74.
[27]吴清平,董晓晖,张菊梅,等.阪崎肠杆菌分类与致病机制[J].微生物学报. 2010(7): 841-846.
[28] Leclercq A, Wanegue C, Baylac P. Comparison of fecal coliform agar and violet red bile lactose agar for fecal coliform enumeration in foods[J]. Appl Environ Microbiol. 2002, 68(4): 1631-1638.
[29]裴晓燕,刘秀梅.中国市售配方粉中阪崎肠杆菌和其它肠杆菌的污染状况[J].中国食品学报. 2006(5): 6-10.
[30] Friedemann M. Enterobacter sakazakii in food and beverages (other than infant formula and milk powder)[J]. Int J Food Microbiol. 2007, 116(1): 1-10.
[31] Kandhai M C, Reij M W, Gorris L G, et al. Occurrence of Enterobacter sakazakii in food production environments and households[J]. Lancet. 2004, 363(9402): 39-40.
[32] Kim H, Beuchat L R. Survival and growth of Enterobacter sakazakii on fresh-cut fruits and vegetables and in unpasteurized juices as affected by storage temperature[J]. J Food Prot. 2005, 68(12): 2541-2552.
[33]袁永辉.阪崎肠杆菌——食品安全控制新目标[J].中国检验检疫. 2005(3): 62.
[34]高涛.食品中金黄色葡萄球菌肠毒素及检测方法的研究进展[J].福建分析测试. 2003(2): 39-42.
[35]张严峻,王志刚,程苏云,等.金黄色葡萄球菌食品和病人分离株肠毒素基因和耐药性比较[J].中国卫生检验杂志. 2008(1): 33-35.
[36] Hiramatsu K, Okuma K, Ma X X, et al. New trends in Staphylococcus aureus infections: glycopeptide resistance in hospital and methicillin resistance in the community[J]. Curr Opin Infect Dis. 2002, 15(4): 407-413.
[37]张红,罗金沪,乐嘉良.一起学校食堂金黄色葡萄球菌食物中毒的调查[J].海峡预防医学杂志. 2005(1): 57.
[38]方月华,陈穗芬,邓志豪.一起金黄色葡萄球菌引起食物中毒调查[J].中国公共卫生. 2006(8): 1004.
[39]刘川玲.一例金黄色葡萄球菌食物中毒报告[J].安徽预防医学杂志. 2008(1): 53.
[40]王国荣,蔡利民.一起金黄色葡萄球菌食物中毒调查[J].浙江预防医学. 2008(12): 37-42.
[41]钟天辉,刘怀志,张辉,等.一起金黄色葡萄球菌食物中毒调查[J].预防医学情报杂志. 2011(9): 735-736.
[42] Arvidson S, Tegmark K. Regulation of virulence determinants in Staphylococcus aureus[J]. Int J Med Microbiol. 2001, 291(2): 159-170.
[43] Zschock M, Kloppert B, Wolter W, et al. Pattern of enterotoxin genes seg, seh, sei and sej positiveStaphylococcus aureus isolated from bovine mastitis[J]. Vet Microbiol. 2005, 108(3-4): 243-249.
[44] Lovseth A, Loncarevic S, Berdal K G. Modified multiplex PCR method for detection of pyrogenic exotoxin genes in staphylococcal isolates[J]. J Clin Microbiol. 2004, 42(8): 3869-3872.
[45]王韶,刘桂华,乔凤,等.食品中金黄色葡萄球菌的PCR检测[J].吉林大学学报(医学版). 2006(5): 933-936.
[46]巢国祥,焦新安,周丽萍,等.食源性金黄色葡萄球菌流行特征、产肠毒素特性及耐药性研究[J].中国卫生检验杂志. 2006(8): 904-907.
[47]邓泽静,李毅,洪程基,等.食品中金黄色葡萄球菌污染状况及其毒素检测[J].浙江预防医学. 2011(2): 92-93.
[48]刘云国.食品卫生微生物学标准鉴定图谱[M].北京:科学教育出版社, 2009.
[49]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社, 2001, 246-247.
[50]伯杰细菌鉴定手册第八版,中译本,科学出版社, 1984年, 729-795.
[51] Stenfors A L, Fagerlund A, Granum P E. From soil to gut: Bacillus cereus and its food poisoning toxins[J]. FEMS Microbiol Rev. 2008, 32(4): 579-606.
[52]齐桂凤.一次由蜡样芽胞杆菌引起的列车食物中毒[J].肉品卫生. 1996(7): 15-17.
[53]王春.蜡样芽胞杆菌引起呼吸道感染1例[J].中华医院感染学杂志. 2003(1): 74.
[54]周帼萍,梁天光,丁淑娟. 1986-2007年中国299起蜡样芽胞杆菌食物中毒案例分析[J].中国食品卫生杂志. 2009(5): 450-454.
[55]邓怀云.蜡样芽胞杆菌食物中毒死亡1例[J].法医学杂志. 1995(3): 137.
[56]唐洪.吃字好担忧——饮服市场假冒伪劣忧思录[J].中国技术监督. 1995(4): 17-19.
[57]李洪春,黄家兴.蜡样芽胞杆菌败血症死亡二例[J].中华医学检验杂志. 1997(1): 57.
[58]翟娅,周运书.一起蜡样芽胞杆菌致家庭食物中毒的调查[J].现代预防医学. 2000(2): 248.
[59]丁国盛,刘良,彭东兵.蜡样芽孢杆菌性食物中毒2例[J].刑事技术. 2000(3): 44.
[60]魏洪贵,刘思寨,刘祥俐,等.腊样芽孢杆菌引起食物中毒的调查分析[J].预防医学情报杂志. 2000(3): 258-259.
[61]许斌,付裕,张革辉.一起食物中毒死亡报告分析[J].实用预防医学. 2003(5): 779.
[62] Stenfors A L, Fagerlund A, Granum P E. From soil to gut: Bacillus cereus and its food poisoning toxins[J]. FEMS Microbiol Rev. 2008, 32(4): 579-606.
[63] Granum P E, Lund T. Bacillus cereus and its food poisoning toxins[J]. FEMS Microbiol Lett. 1997, 157(2): 223-228.
[64] Kreske A C, Ryu J H, Pettigrew C A, et al. Lethality of chlorine, chlorine dioxide, and a commercial produce sanitizer to Bacillus cereus and Pseudomonas in a liquid detergent, on stainless steel, and in biofilm[J]. J Food Prot. 2006, 69(11): 2621-2634.
[65]赵文彬,周克捷,李家富,等.应用免疫磁珠法分离大肠杆菌O_(157)H_7[J].现代预防医学. 2000(3): 331-332.
[66] Lynch M J, Leon-Velarde C G, Mcewen S, et al. Evaluation of an automated immunomagnetic separation method for the rapid detection of Salmonella species in poultry environmental samples[J]. J Microbiol Methods. 2004, 58(2): 285-288.
[67] Inat G, Siriken B. Detection of Escherichia coli O157 and Escherichia coli O157:H7 by the immunomagnetic separation technique and stx1 and stx2 genes by multiplex PCR in slaughtered cattle in Samsun Province, Turkey[J]. J Vet Sci. 2010, 11(4): 321-326.
[68]王小红,徐康,王莹,等.金黄色葡萄球菌B型肠毒素间接竞争ELISA检测方法的建立[J].食品科学. 2006(8): 227-231.
[69]黄素珍,杜元钊,张艳红.检测肠炎沙门氏菌ELISA方法的建立与应用研究[J].中国预防兽医学报. 2006(2): 196-200.
[70] Jiao Y, Zeng X, Guo X, et al. Preparation and evaluation of recombinant severe fever with thrombocytopenia syndrome virus nucleocapsid protein for detection of total antibodies in human and animal sera by double-antigen sandwich enzyme-linked immunosorbent assay[J]. J Clin Microbiol. 2012, 50(2): 372-377.
[71]宋农,李君文,王新为,等.胶体金免疫层析法用于产肠毒素葡萄球菌快速检测的研究[J].中华检验医学杂志. 2003(7): 54-55.
[72]高成秀,孙建和,严亚贤.大肠杆菌O_(157)及其Stx2免疫胶体金层析检测方法的建立[J].中国兽医科学. 2009(2): 154-158.
[73] Jung B Y, Jung S C, Kweon C H. Development of a rapid immunochromatographic strip for detection of Escherichia coli O157[J]. J Food Prot. 2005, 68(10): 2140-2143.
[74] Rasooly A, Rasooly R S. Detection and analysis of Staphylococcal enterotoxin A in food by Western immunoblotting[J]. Int J Food Microbiol. 1998, 41(3): 205-212.
[75]陈海英,番敏,张柳.免疫印迹技术在临床幽门螺杆菌检测中的应用[J].中国实验诊断学. 2006(9): 995-996.
[76] Sharma N K, Rees C E, Dodd C E. Development of a single-reaction multiplex PCR toxin typing assay for Staphylococcus aureus strains[J]. Appl Environ Microbiol. 2000, 66(4): 1347-1353.
[77]黄留玉. PCR最新技术原理、方法及应用[R].湖北:湖北人民出版社, 2006.
[78]胡慧,段志刚,崔保安,等.动物源大肠埃希菌O157∶H7多重PCR检测方法的初步研究[J].西北农林科技大学学报(自然科学版). 2011(3): 34-39.
[79]刘继超,姜铁民,陈历俊,等.多重PCR检测原料乳中沙门氏菌、无乳链球菌和金黄色葡萄球菌的研究[J].中国食品学报. 2010(6): 173-179.
[80]彭雁忠,贾宏,邹虹,等.多联实时荧光PCR定量方法检测四种肠道细菌的研究[J].中国热带医学. 2005(5): 943-947.
[81]张朝.荧光定量PCR检测婴儿配方奶粉中阪崎肠杆菌研究[D].河北:河北农业大学, 2007.
[82]李一松,韩希妍,赵凤,等. Taqman探针实时PCR检测金黄色葡萄球菌的研究[J].食品与发酵工业. 2008(5): 156-161.
[83] Walker G T, Little M C, Nadeau J G, et al. Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system[J]. Proc Natl Acad Sci U S A. 1992, 89(1): 392-396.
[84]梁炜,尼秀媚,雷质文,等.霍乱弧菌SDA-琼脂糖凝胶电泳检测技术研究[J].食品安全质量检测学报. 2011(3): 165-169.
[85] Kanayama A, Fujihara E, Saika T, et al. [Detection of Chlamydia trachomatis and Neisseriagonorrhoeae in urine samples of males and females by the strand displacement amplification (SDA) method][J]. Kansenshogaku Zasshi. 2008, 82(3): 182-186.
[86] Nuovo G J. In situ strand displacement amplification: an improved technique for the detection of low copy nucleic acids[J]. Diagn Mol Pathol. 2000, 9(4): 195-202.
[87] Vincent M, Xu Y, Kong H. Helicase-dependent isothermal DNA amplification[J]. EMBO Rep. 2004, 5(8): 795-800.
[88] Gill P, Amini M, Ghaemi A, et al. Detection of Helicobacter pylori by enzyme-linked immunosorbent assay of thermophilic helicase-dependent isothermal DNA amplification[J]. Diagn Microbiol Infect Dis. 2007, 59(3): 243-249.
[89]王建广,雷质文,石琰璟,等.大肠埃希氏菌O157:H7的HDA检测方法的建立[J].安徽农业大学学报. 2011(2): 271-274.
[90] Goldmeyer J, Kong H, Tang W. Development of a novel one-tube isothermal reverse transcription thermophilic helicase-dependent amplification platform for rapid RNA detection[J]. J Mol Diagn. 2007, 9(5): 639-644.
[91] Notomi T, Okayama H, Masubuchi H, et al. Loop-mediated isothermal amplification of DNA[J]. Nucleic Acids Res. 2000, 28(12): E63.
[92]贺楠,雷质文,高宏伟,等.阪崎肠杆菌环介导恒温扩增方法检测[J].中国公共卫生. 2009(4): 509-511.
[93]高宏伟,徐彪,朱来华,等.应用LAMP检测方法检测肉制品中的单增李斯特菌[J].食品安全质量检测学报. 2010(1): 12-17.
[94] Song T, Toma C, Nakasone N, et al. Sensitive and rapid detection of Shigella and enteroinvasive Escherichia coli by a loop-mediated isothermal amplification method[J]. FEMS Microbiol Lett. 2005, 243(1): 259-263.
[95] Tang Y, Diao Y, Yu C, et al. Rapid Detection of Tembusu Virus by Reverse-Transcription, Loop-mediated Isothermal Amplification (RT-LAMP)[J]. Transbound Emerg Dis. 2011.
[96]黄冠军,殷幼平,张仑,等.柑桔溃疡病菌滚环扩增检测体系的建立[J].微生物学报. 2008(3): 375-379.
[97]焦肖霞.利用滚环扩增反应检测端粒酶活性及单核苷酸多态性[D].河北:河北大学, 2010.
[98] Whitworth K M, Agca C, Kim J G, et al. Transcriptional profiling of pig embryogenesis by using a 15-K member unigene set specific for pig reproductive tissues and embryos[J]. Biol Reprod. 2005, 72(6): 1437-1451.
[99] Lau L T, Reid S M, King D P, et al. Detection of foot-and-mouth disease virus by nucleic acid sequence-based amplification (NASBA)[J]. Vet Microbiol. 2008, 126(1-3): 101-110.
[100]张志宏,李丽丽,杨洪一.利用NASBA技术检测草莓斑驳病毒[J].果树学报. 2007(6): 858-862.
[101]薛力刚,刘金华,王全凯,等.金黄葡萄球菌核酸探针检测方法的建立[J].中国生物制品学杂志. 2010(1): 91-94.李瑶.基因芯片与功能基因组[M].北京:化学工业出版社, 2004.
[102]李瑶.基因芯片与功能基因组[M].北京:化学工业出版社, 2004.
[103]聂萌.通用芯片技术检测和鉴定食源致病菌方法的研究[D].陕西:西北大学, 2006.
[104] Chizhikov V, Rasooly A, Chumakov K, et al. Microarray analysis of microbial virulence factors[J]. Appl Environ Microbiol. 2001, 67(7): 3258-3263.
[105] Homola J, Dostalek J, Chen S, et al. Spectral surface plasmon resonance biosensor for detection of staphylococcal enterotoxin B in milk[J]. Int J Food Microbiol. 2002, 75(1-2): 61-69.
[106] Geng T, Morgan M T, Bhunia A K. Detection of low levels of Listeria monocytogenes cells by using a fiber-optic immunosensor[J]. Appl Environ Microbiol. 2004, 70(10): 6138-6146.
[107] Mohan N 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.
[108]刘书亮,刘冬香,贾仁勇,等.多重PCR检测食源金黄色葡萄球菌nuc、blaZ和mecA基因方法的建立与应用[J].中国人兽共患病学报. 2010(5): 475-480.
[109]齐哲,张伟,刘卫华,等. FTA滤膜与环介导等温扩增技术结合快速检测消毒乳中的蜡样芽孢杆菌[J].中国食品学报. 2009(3): 156-161.
[2]刘秀梅.食源性疾病仍是中国头号食品安全问题[R].北京:中国疾病预防控制中心, 2006.
[3]汪清,葛秀清,黄祖华.细菌性食物中毒的现状及检验[J].中国动物检疫. 2004(4): 20-22.
[4]陈建琳,刘明辉.细菌性食物中毒流行趋势及预防对策[J].中国卫生检验杂志. 2002(4): 481.
[5]邱文毅,钱进,胡月珍,等.食源性疾病现状及国际航行船舶食品卫生监管建议[J].口岸卫生控制. 2011(1): 43-46.
[6]乔铁军,张国祥,王幼雄.食源性疾病及预防探讨[J].浙江预防医学. 2007(11): 35-39.
[7] Mead P S, Slutsker L, Dietz V, et al. Food-related illness and death in the United States[J]. Emerg Infect Dis. 1999, 5(5): 607-625.
[8] Schraff RL. Health-related costs from foodborne illness in the United States.Produce Safety Project [R]. Georgetown University, Washington. D.C. 2010.
[9] Mccabe-Sellers B J, Beattie S E. Food safety: emerging trends in foodborne illness surveillance and prevention[J]. J Am Diet Assoc. 2004, 104(11): 1708-1717.
[10]李建科,陈锦屏.我国食品安全现状与入世后的形势与对策[J].食品科学. 2003(8): 272-276.
[11]王艳君.冷却肉中致病菌的多重PCR检测技术研究[D].河南:河南农业大学, 2007.
[12]吴雅儿.因喂服染有蜡样芽胞杆菌奶粉引起婴儿腹泻1例[J].海峡预防医学杂志. 2003(5): 4.
[13]萨日娜.一起由金黄色葡萄球菌引起的奶粉食物中毒[J].中国卫生检验杂志. 2005(5): 636.
[14]叶红萍.从雅士利婴儿配方奶粉中检出蜡样芽胞杆菌[J].海峡预防医学杂志. 2008(1): 4.
[15] Lehner A, Stephan R. Microbiological, epidemiological, and food safety aspects of Enterobacter sakazakii[J]. J Food Prot. 2004, 67(12): 2850-2857.
[16] Nazarowec-White M, Farber J M. Enterobacter sakazakii: a review[J]. Int J Food Microbiol. 1997, 34(2): 103-113.
[17] Breeuwer P, Lardeau A, Peterz M, et al. Desiccation and heat tolerance of Enterobacter sakazakii[J]. J Appl Microbiol. 2003, 95(5): 967-973.
[18] Iversen C, Druggan P, Forsythe S. A selective differential medium for Enterobacter sakazakii, a preliminary study[J]. Int J Food Microbiol. 2004, 96(2): 133-139.
[19]张爱霞,生庆海,张涛.食品中阪崎肠杆菌分析及其检测[J].中国乳品工业. 2005(10): 43-45.
[20] Muytjens H L, Zanen H C, Sonderkamp H J, et al. Analysis of eight cases of neonatal meningitis and sepsis due to Enterobacter sakazakii[J]. J Clin Microbiol. 1983, 18(1): 115-120.
[21]徐英春,张小江,陈民钧,等.肠杆菌属的耐药调查及抗感染用药探讨[J].中华医院感染学杂志. 2001(3): 77-79.
[22] Enterobacter sakazakii infections associated with the use of powdered infant formula--Tennessee, 2001[J]. MMWR Morb Mortal Wkly Rep. 2002, 51(14): 297-300.
[23] Bowen A B, Braden C R. Invasive Enterobacter sakazakii disease in infants[J]. Emerg Infect Dis. 2006, 12(8): 1185-1189.
[24] Teramoto S, Tanabe Y, Okano E, et al. A first fatal neonatal case of Enterobacter sakazakii infection in Japan[J]. Pediatr Int. 2010, 52(2): 312-313.
[25] Pagotto F J, Nazarowec-White M, Bidawid S, et al. Enterobacter sakazakii: infectivity and enterotoxin production in vitro and in vivo[J]. J Food Prot. 2003, 66(3): 370-375.
[26] Townsend S, Caubilla B J, Loc-Carrillo C, et al. The presence of endotoxin in powdered infant formula milk and the influence of endotoxin and Enterobacter sakazakii on bacterial translocation in the infant rat[J]. Food Microbiol. 2007, 24(1): 67-74.
[27]吴清平,董晓晖,张菊梅,等.阪崎肠杆菌分类与致病机制[J].微生物学报. 2010(7): 841-846.
[28] Leclercq A, Wanegue C, Baylac P. Comparison of fecal coliform agar and violet red bile lactose agar for fecal coliform enumeration in foods[J]. Appl Environ Microbiol. 2002, 68(4): 1631-1638.
[29]裴晓燕,刘秀梅.中国市售配方粉中阪崎肠杆菌和其它肠杆菌的污染状况[J].中国食品学报. 2006(5): 6-10.
[30] Friedemann M. Enterobacter sakazakii in food and beverages (other than infant formula and milk powder)[J]. Int J Food Microbiol. 2007, 116(1): 1-10.
[31] Kandhai M C, Reij M W, Gorris L G, et al. Occurrence of Enterobacter sakazakii in food production environments and households[J]. Lancet. 2004, 363(9402): 39-40.
[32] Kim H, Beuchat L R. Survival and growth of Enterobacter sakazakii on fresh-cut fruits and vegetables and in unpasteurized juices as affected by storage temperature[J]. J Food Prot. 2005, 68(12): 2541-2552.
[33]袁永辉.阪崎肠杆菌——食品安全控制新目标[J].中国检验检疫. 2005(3): 62.
[34]高涛.食品中金黄色葡萄球菌肠毒素及检测方法的研究进展[J].福建分析测试. 2003(2): 39-42.
[35]张严峻,王志刚,程苏云,等.金黄色葡萄球菌食品和病人分离株肠毒素基因和耐药性比较[J].中国卫生检验杂志. 2008(1): 33-35.
[36] Hiramatsu K, Okuma K, Ma X X, et al. New trends in Staphylococcus aureus infections: glycopeptide resistance in hospital and methicillin resistance in the community[J]. Curr Opin Infect Dis. 2002, 15(4): 407-413.
[37]张红,罗金沪,乐嘉良.一起学校食堂金黄色葡萄球菌食物中毒的调查[J].海峡预防医学杂志. 2005(1): 57.
[38]方月华,陈穗芬,邓志豪.一起金黄色葡萄球菌引起食物中毒调查[J].中国公共卫生. 2006(8): 1004.
[39]刘川玲.一例金黄色葡萄球菌食物中毒报告[J].安徽预防医学杂志. 2008(1): 53.
[40]王国荣,蔡利民.一起金黄色葡萄球菌食物中毒调查[J].浙江预防医学. 2008(12): 37-42.
[41]钟天辉,刘怀志,张辉,等.一起金黄色葡萄球菌食物中毒调查[J].预防医学情报杂志. 2011(9): 735-736.
[42] Arvidson S, Tegmark K. Regulation of virulence determinants in Staphylococcus aureus[J]. Int J Med Microbiol. 2001, 291(2): 159-170.
[43] Zschock M, Kloppert B, Wolter W, et al. Pattern of enterotoxin genes seg, seh, sei and sej positiveStaphylococcus aureus isolated from bovine mastitis[J]. Vet Microbiol. 2005, 108(3-4): 243-249.
[44] Lovseth A, Loncarevic S, Berdal K G. Modified multiplex PCR method for detection of pyrogenic exotoxin genes in staphylococcal isolates[J]. J Clin Microbiol. 2004, 42(8): 3869-3872.
[45]王韶,刘桂华,乔凤,等.食品中金黄色葡萄球菌的PCR检测[J].吉林大学学报(医学版). 2006(5): 933-936.
[46]巢国祥,焦新安,周丽萍,等.食源性金黄色葡萄球菌流行特征、产肠毒素特性及耐药性研究[J].中国卫生检验杂志. 2006(8): 904-907.
[47]邓泽静,李毅,洪程基,等.食品中金黄色葡萄球菌污染状况及其毒素检测[J].浙江预防医学. 2011(2): 92-93.
[48]刘云国.食品卫生微生物学标准鉴定图谱[M].北京:科学教育出版社, 2009.
[49]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社, 2001, 246-247.
[50]伯杰细菌鉴定手册第八版,中译本,科学出版社, 1984年, 729-795.
[51] Stenfors A L, Fagerlund A, Granum P E. From soil to gut: Bacillus cereus and its food poisoning toxins[J]. FEMS Microbiol Rev. 2008, 32(4): 579-606.
[52]齐桂凤.一次由蜡样芽胞杆菌引起的列车食物中毒[J].肉品卫生. 1996(7): 15-17.
[53]王春.蜡样芽胞杆菌引起呼吸道感染1例[J].中华医院感染学杂志. 2003(1): 74.
[54]周帼萍,梁天光,丁淑娟. 1986-2007年中国299起蜡样芽胞杆菌食物中毒案例分析[J].中国食品卫生杂志. 2009(5): 450-454.
[55]邓怀云.蜡样芽胞杆菌食物中毒死亡1例[J].法医学杂志. 1995(3): 137.
[56]唐洪.吃字好担忧——饮服市场假冒伪劣忧思录[J].中国技术监督. 1995(4): 17-19.
[57]李洪春,黄家兴.蜡样芽胞杆菌败血症死亡二例[J].中华医学检验杂志. 1997(1): 57.
[58]翟娅,周运书.一起蜡样芽胞杆菌致家庭食物中毒的调查[J].现代预防医学. 2000(2): 248.
[59]丁国盛,刘良,彭东兵.蜡样芽孢杆菌性食物中毒2例[J].刑事技术. 2000(3): 44.
[60]魏洪贵,刘思寨,刘祥俐,等.腊样芽孢杆菌引起食物中毒的调查分析[J].预防医学情报杂志. 2000(3): 258-259.
[61]许斌,付裕,张革辉.一起食物中毒死亡报告分析[J].实用预防医学. 2003(5): 779.
[62] Stenfors A L, Fagerlund A, Granum P E. From soil to gut: Bacillus cereus and its food poisoning toxins[J]. FEMS Microbiol Rev. 2008, 32(4): 579-606.
[63] Granum P E, Lund T. Bacillus cereus and its food poisoning toxins[J]. FEMS Microbiol Lett. 1997, 157(2): 223-228.
[64] Kreske A C, Ryu J H, Pettigrew C A, et al. Lethality of chlorine, chlorine dioxide, and a commercial produce sanitizer to Bacillus cereus and Pseudomonas in a liquid detergent, on stainless steel, and in biofilm[J]. J Food Prot. 2006, 69(11): 2621-2634.
[65]赵文彬,周克捷,李家富,等.应用免疫磁珠法分离大肠杆菌O_(157)H_7[J].现代预防医学. 2000(3): 331-332.
[66] Lynch M J, Leon-Velarde C G, Mcewen S, et al. Evaluation of an automated immunomagnetic separation method for the rapid detection of Salmonella species in poultry environmental samples[J]. J Microbiol Methods. 2004, 58(2): 285-288.
[67] Inat G, Siriken B. Detection of Escherichia coli O157 and Escherichia coli O157:H7 by the immunomagnetic separation technique and stx1 and stx2 genes by multiplex PCR in slaughtered cattle in Samsun Province, Turkey[J]. J Vet Sci. 2010, 11(4): 321-326.
[68]王小红,徐康,王莹,等.金黄色葡萄球菌B型肠毒素间接竞争ELISA检测方法的建立[J].食品科学. 2006(8): 227-231.
[69]黄素珍,杜元钊,张艳红.检测肠炎沙门氏菌ELISA方法的建立与应用研究[J].中国预防兽医学报. 2006(2): 196-200.
[70] Jiao Y, Zeng X, Guo X, et al. Preparation and evaluation of recombinant severe fever with thrombocytopenia syndrome virus nucleocapsid protein for detection of total antibodies in human and animal sera by double-antigen sandwich enzyme-linked immunosorbent assay[J]. J Clin Microbiol. 2012, 50(2): 372-377.
[71]宋农,李君文,王新为,等.胶体金免疫层析法用于产肠毒素葡萄球菌快速检测的研究[J].中华检验医学杂志. 2003(7): 54-55.
[72]高成秀,孙建和,严亚贤.大肠杆菌O_(157)及其Stx2免疫胶体金层析检测方法的建立[J].中国兽医科学. 2009(2): 154-158.
[73] Jung B Y, Jung S C, Kweon C H. Development of a rapid immunochromatographic strip for detection of Escherichia coli O157[J]. J Food Prot. 2005, 68(10): 2140-2143.
[74] Rasooly A, Rasooly R S. Detection and analysis of Staphylococcal enterotoxin A in food by Western immunoblotting[J]. Int J Food Microbiol. 1998, 41(3): 205-212.
[75]陈海英,番敏,张柳.免疫印迹技术在临床幽门螺杆菌检测中的应用[J].中国实验诊断学. 2006(9): 995-996.
[76] Sharma N K, Rees C E, Dodd C E. Development of a single-reaction multiplex PCR toxin typing assay for Staphylococcus aureus strains[J]. Appl Environ Microbiol. 2000, 66(4): 1347-1353.
[77]黄留玉. PCR最新技术原理、方法及应用[R].湖北:湖北人民出版社, 2006.
[78]胡慧,段志刚,崔保安,等.动物源大肠埃希菌O157∶H7多重PCR检测方法的初步研究[J].西北农林科技大学学报(自然科学版). 2011(3): 34-39.
[79]刘继超,姜铁民,陈历俊,等.多重PCR检测原料乳中沙门氏菌、无乳链球菌和金黄色葡萄球菌的研究[J].中国食品学报. 2010(6): 173-179.
[80]彭雁忠,贾宏,邹虹,等.多联实时荧光PCR定量方法检测四种肠道细菌的研究[J].中国热带医学. 2005(5): 943-947.
[81]张朝.荧光定量PCR检测婴儿配方奶粉中阪崎肠杆菌研究[D].河北:河北农业大学, 2007.
[82]李一松,韩希妍,赵凤,等. Taqman探针实时PCR检测金黄色葡萄球菌的研究[J].食品与发酵工业. 2008(5): 156-161.
[83] Walker G T, Little M C, Nadeau J G, et al. Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system[J]. Proc Natl Acad Sci U S A. 1992, 89(1): 392-396.
[84]梁炜,尼秀媚,雷质文,等.霍乱弧菌SDA-琼脂糖凝胶电泳检测技术研究[J].食品安全质量检测学报. 2011(3): 165-169.
[85] Kanayama A, Fujihara E, Saika T, et al. [Detection of Chlamydia trachomatis and Neisseriagonorrhoeae in urine samples of males and females by the strand displacement amplification (SDA) method][J]. Kansenshogaku Zasshi. 2008, 82(3): 182-186.
[86] Nuovo G J. In situ strand displacement amplification: an improved technique for the detection of low copy nucleic acids[J]. Diagn Mol Pathol. 2000, 9(4): 195-202.
[87] Vincent M, Xu Y, Kong H. Helicase-dependent isothermal DNA amplification[J]. EMBO Rep. 2004, 5(8): 795-800.
[88] Gill P, Amini M, Ghaemi A, et al. Detection of Helicobacter pylori by enzyme-linked immunosorbent assay of thermophilic helicase-dependent isothermal DNA amplification[J]. Diagn Microbiol Infect Dis. 2007, 59(3): 243-249.
[89]王建广,雷质文,石琰璟,等.大肠埃希氏菌O157:H7的HDA检测方法的建立[J].安徽农业大学学报. 2011(2): 271-274.
[90] Goldmeyer J, Kong H, Tang W. Development of a novel one-tube isothermal reverse transcription thermophilic helicase-dependent amplification platform for rapid RNA detection[J]. J Mol Diagn. 2007, 9(5): 639-644.
[91] Notomi T, Okayama H, Masubuchi H, et al. Loop-mediated isothermal amplification of DNA[J]. Nucleic Acids Res. 2000, 28(12): E63.
[92]贺楠,雷质文,高宏伟,等.阪崎肠杆菌环介导恒温扩增方法检测[J].中国公共卫生. 2009(4): 509-511.
[93]高宏伟,徐彪,朱来华,等.应用LAMP检测方法检测肉制品中的单增李斯特菌[J].食品安全质量检测学报. 2010(1): 12-17.
[94] Song T, Toma C, Nakasone N, et al. Sensitive and rapid detection of Shigella and enteroinvasive Escherichia coli by a loop-mediated isothermal amplification method[J]. FEMS Microbiol Lett. 2005, 243(1): 259-263.
[95] Tang Y, Diao Y, Yu C, et al. Rapid Detection of Tembusu Virus by Reverse-Transcription, Loop-mediated Isothermal Amplification (RT-LAMP)[J]. Transbound Emerg Dis. 2011.
[96]黄冠军,殷幼平,张仑,等.柑桔溃疡病菌滚环扩增检测体系的建立[J].微生物学报. 2008(3): 375-379.
[97]焦肖霞.利用滚环扩增反应检测端粒酶活性及单核苷酸多态性[D].河北:河北大学, 2010.
[98] Whitworth K M, Agca C, Kim J G, et al. Transcriptional profiling of pig embryogenesis by using a 15-K member unigene set specific for pig reproductive tissues and embryos[J]. Biol Reprod. 2005, 72(6): 1437-1451.
[99] Lau L T, Reid S M, King D P, et al. Detection of foot-and-mouth disease virus by nucleic acid sequence-based amplification (NASBA)[J]. Vet Microbiol. 2008, 126(1-3): 101-110.
[100]张志宏,李丽丽,杨洪一.利用NASBA技术检测草莓斑驳病毒[J].果树学报. 2007(6): 858-862.
[101]薛力刚,刘金华,王全凯,等.金黄葡萄球菌核酸探针检测方法的建立[J].中国生物制品学杂志. 2010(1): 91-94.李瑶.基因芯片与功能基因组[M].北京:化学工业出版社, 2004.
[102]李瑶.基因芯片与功能基因组[M].北京:化学工业出版社, 2004.
[103]聂萌.通用芯片技术检测和鉴定食源致病菌方法的研究[D].陕西:西北大学, 2006.
[104] Chizhikov V, Rasooly A, Chumakov K, et al. Microarray analysis of microbial virulence factors[J]. Appl Environ Microbiol. 2001, 67(7): 3258-3263.
[105] Homola J, Dostalek J, Chen S, et al. Spectral surface plasmon resonance biosensor for detection of staphylococcal enterotoxin B in milk[J]. Int J Food Microbiol. 2002, 75(1-2): 61-69.
[106] Geng T, Morgan M T, Bhunia A K. Detection of low levels of Listeria monocytogenes cells by using a fiber-optic immunosensor[J]. Appl Environ Microbiol. 2004, 70(10): 6138-6146.
[107] Mohan N 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.
[108]刘书亮,刘冬香,贾仁勇,等.多重PCR检测食源金黄色葡萄球菌nuc、blaZ和mecA基因方法的建立与应用[J].中国人兽共患病学报. 2010(5): 475-480.
[109]齐哲,张伟,刘卫华,等. FTA滤膜与环介导等温扩增技术结合快速检测消毒乳中的蜡样芽孢杆菌[J].中国食品学报. 2009(3): 156-161.