AN IMPROVED ASSAY FOR IDENTIFICATION of TYPE A CLOSTRIDIUM BOTULINUM USING the POLYMERASE CHAIN REACTION1
作者:
JOSEPH L. FERREIRA,
MOSTAFA K. HAMDY,
STEVEN G. MCCAY,
BARBARA R. BAUMSTARK,
期刊:
Journal of Rapid Methods&Automation in Microbiology
(WILEY Available online 1992)
卷期:
Volume 1,
issue 1
页码: 29-39
ISSN:1060-3999
年代: 1992
DOI:10.1111/j.1745-4581.1992.tb00068.x
出版商: Blackwell Publishing Ltd
数据来源: WILEY
摘要:
The polymerase chain reaction (PCR) was used to amplify a 1340‐bp fragment from the toxigenicClostridium botulinumtype A chromosome. Two oligonucleotide primers were synthesized to bracket the sequence coding for most of the type A toxin light chain (1340‐bp in size). the PCR amplified a 1340‐bp fragment from the DNA of all elevenC. botulinumtype A strains tested. In contrast, no DNA fragments were amplified fromC. botulinumtypes B, E, and F or from the other Clostridial chromosomal DNA examined. the products amplifed from botulinal type A strains were digested to produce fragments of the size predicted from the known location of theHinddIII sites within the toxin gene sequence. When used as a DNA probe, the 1340‐bp PCR generated fragment from type A C.botulinumstrain 73A hybridized to the PCR products from all 11 type A botulinal strains and to type A thromosomal DNA prepa
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