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细菌ATP荧光检测与常规计数相关性研究

时间:2015-03-30 12:17 文章来源:http://www.lunwenbuluo.com 作者:李佳明等 点击次数:

 摘要: ATP荧光检测仪是近年来的新生产物,可以通过荧光值快速反映出细菌菌落数。本研究以金黄色葡萄球菌(ATCC 6538) 、大肠杆菌(8099)为试材,培养24 h后,各自稀释11个梯度(10倍稀释),并进行细菌计数;同时采用ATP荧光仪对每一个稀释梯度进行荧光检测。以lg(菌落数)为横坐标(x),以ATP荧光值(y)为纵坐标,分别进行线性、对数、乘幂、指数拟合,发现二者之间关系符合指数曲线,相关系数均接近0.9或以上。以屠宰场的要求“<1 000荧光值为合格,>2 000荧光值为不合格”为实例进行使用指导:当实际lg(菌落数)>10.04时,得出不合格的结论;当实际lg(菌落数)<8.27时,得出合格毕业论文格式的结论;但如果8.27<实际lg(菌落数)<10.04时,则容易出现误判。结论认为ATP荧光检测可以在一定程度上代替细菌常规检测,并应用于实际生产。 

  关键词:ATP荧光检测仪;细菌;常规计数;相关性 
  中图分类号:Q93-331文献标识号:A文章编号:1001-4942(2015)02-0111-04 
  AbstractATP fluorescence detector, as a new technical device, can rapidly reflect the amounts of bacterial colonies via fluorescence value. Selecting Staphylococcus aureus ATCC 6538 and Escherichia coli 8099 as materials, the number of colonies were counted after cultured for 24 hours and diluted to 11 gradients (10 times dilution). Meanwhile, the fluorescence value of each dilution was detected by ATP fluorescence detector. On the basis of these results, with lg(CFU) as abscissa (x) and ATP fluorescence value as ordinate (y), the linear fitting, logarithm fitting and exponent fitting were conducted;it was found that the relationship between lg(CFU) and ATP fluorescence value was in accordance with exponential curve with the correlation coefficient of close to 0.9 or above. According to the requirement of slaughterhouse, if the fluorescence value was less than 1 000, it is qualified; if fluorescence value was greater than 2 000, it is not qualified. It could be explained as follows. If actual lg(CFU) was greater than 10.04, it should be unqualified; if actual lg (CFU) was less than 8.27, it should be qualified. But when actual lg(CFU) was between 8.27 and 10.04, it should be prone to misjudge. Ultimately, to some extent, the ATP fluorescence detection was able to replace the bacteria conventional detection to be used in actual production. 
  Key wordsATP fluorescence detector; Bacteria; Conventional count; Correlation 
  在细菌总数的检测中,传统的培养法由于检测时间较长,已经不能满足食品企业实施HACCP体系时实时监控的要求1。20世纪80年代,英国首先研制出ATP 检测系统,随后发展到欧洲、美国和日本。应用范围涉及食品加工等行业,检测内容包括生物和食品残渣2;快速检测宇宙空间水系统中微生物量也是该技术较早应用的领域之一3;也有将ATP (三磷酸腺苷)生物荧光法引入到医疗机构器械清洗前、后残留物检测4。在我国,基于国家发明专利“含虫光素基因的新型菌株及虫光素酶的生产方法”5的一种荧光检测仪逐步被应用,其能通过ATP荧光仪与Ultrasnap配合使用,采用生物发光技术检测残留的ATP作为表面清洁度的指标。本研究即通过ATP计数与细菌常规检测两种方法互相印证,探索二者之间的相关性,并举例得出几个节点数据指导实践应用。 
  1材料与方法 
  1.1试剂与设备 
  

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