Quantitative duplex PCR of Clostridium botulinum types A and B neurotoxin genes

被引:28
作者
Kasai, Yoshiaki [1 ]
Kimura, Bon [1 ]
Tajima, Yosuke [1 ]
Fujii, Tateo [1 ]
机构
[1] Tokyo Univ Marine Sci & Technol, Fac Marine Sci, Dept Food Sci & Technol, Minato Ku, Tokyo 1088477, Japan
来源
JOURNAL OF THE FOOD HYGIENIC SOCIETY OF JAPAN | 2007年 / 48卷 / 01期
关键词
Clostridium botulinum; quantitative PCR; duplex; TaqMan; POLYMERASE-CHAIN-REACTION; LINKED-IMMUNOSORBENT-ASSAY; REVERSE TRANSCRIPTION-PCR; REAL-TIME PCR; MOUSE BIOASSAY; SODIUM-NITRITE; FOOD SAMPLES; TOXIN GENES; IDENTIFICATION; EXPRESSION;
D O I
10.3358/shokueishi.48.19
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
A duplex quantitative polymerase chain reaction (PCR) assay for Clostridium botulinum types A and B was developed. The sensitivity and specificity of the assay were verified by using 6 strains of type A, 7 strains of type 13, and 14 genera of 42 non-C. botulinum types A and B strains, including C. botulinum types C, D, E, F, and G. In pure culture, the detection limit was 10(2) CFU/mL for type A and 10(3) CFU/mL for type B. In mushroom broth, increases in the amounts of C. botulinum types A and B could be monitored separately (the quantifiable range was 10(2) to 10(6) for type A and 10(2) to 10(7) for type 13) from each sample that contained a large number of background bacteria, and toxin could be detected much earlier than with mouse assay. These results suggest that duplex quantitative PCR methods are useful to detect and quantify C. botulinum types A and/or B toxin genes.
引用
收藏
页码:19 / 26
页数:8
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