PCR-Based Method Using Propidium Monoazide To Distinguish Viable from Nonviable Bacillus subtilis Spores

被引:74
作者
Rawsthorne, H. [1 ]
Dock, C. N. [1 ]
Jaykus, L. A. [1 ]
机构
[1] N Carolina State Univ, Dept Food Bioproc & Nutr Sci, Raleigh, NC 27695 USA
关键词
REAL-TIME PCR; ETHIDIUM MONOAZIDE; DEAD CELLS; DNA;
D O I
10.1128/AEM.02524-08
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
This paper describes a molecular-based method which is able to discriminate between viable and inactivated Bacillus subtilis spores by utilizing the DNA-intercalating dye propidium monoazide. The approach should be valuable in our attempt to employ molecular methods to streamline the evaluation of process validation using bacterial endospores.
引用
收藏
页码:2936 / 2939
页数:4
相关论文
共 9 条
[1]  
Holdsworth S.D., 1997, THERMAL PROCESSING P, DOI DOI 10.1007/978-3-319-24904-9
[2]   Selective removal of DNA from dead cells of mixed bacterial communities by use of ethidium monoazide [J].
Nocker, A ;
Camper, AK .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2006, 72 (03) :1997-2004
[3]   Ethidium monoazide for DNA-based differentiation of viable and dead bacteria by 5′-nuclease PCR [J].
Nogva, HK ;
Dromtorp, SM ;
Nissen, H ;
Rudi, K .
BIOTECHNIQUES, 2003, 34 (04) :804-+
[4]   THERMAL DESTRUCTION OF CLOSTRIDIUM-BOTULINUM SPORES SUSPENDED IN TOMATO JUICE IN ALUMINUM THERMAL DEATH TIME TUBES [J].
ODLAUG, TE ;
PFLUG, IJ .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1977, 34 (01) :23-29
[5]   Detection of viable and dead Listeria monocytogenes on gouda-like cheeses by real-time PCR [J].
Rudi, K ;
Naterstad, K ;
Dromtorp, SM ;
Holo, H .
LETTERS IN APPLIED MICROBIOLOGY, 2005, 40 (04) :301-306
[6]   Use of ethidium monoazide and PCR in combination for quantification of viable and dead cells in complex samples [J].
Rudi, K ;
Moen, B ;
Dromtorp, SM ;
Holck, AL .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2005, 71 (02) :1018-1024
[7]   Development of a real-time PCR assay targeting the sporulation gene, spo0A, for the enumeration of thermophilic bacilli in milk powder [J].
Rueckert, A ;
Ronimus, RS ;
Morgan, HW .
FOOD MICROBIOLOGY, 2006, 23 (03) :220-230
[8]  
STERN JA, 1954, FOOD TECHNOL-CHICAGO, V8, P139
[9]   Discrimination of viable Vibrio vulnificus cells from dead cells in real-time PCR [J].
Wang, SS ;
Levin, RE .
JOURNAL OF MICROBIOLOGICAL METHODS, 2006, 64 (01) :1-8