PCR-based random amplified polymorphic DNA fingerprinting of Yersinia pseudotuberculosis and its practical applications

Makino, Sou‐ichi; Okada, Yumiko; Matsumoto, Tsutomu; Kaneko, Seiji; Sasakawa, Chihiro

doi:10.1128/jcm.32.1.65-69.1994

Cited by 54 publications

(14 citation statements)

References 21 publications

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“…The concentration of MgCl 2 used in the reaction buffer was 1.5 mM, and this concentration enhanced the specificity of the PCR and produced informative arrays in this study (2). In the first round of screening, amplification by 19 of the primers resulted in several clear DNA bands in both strains (data not shown).…”

Section: Resultsmentioning

confidence: 70%

Molecular Typing of Vibrio parahaemolyticus Isolates, Obtained from Patients Involved in Food Poisoning Outbreaks in Taiwan, by Random Amplified Polymorphic DNA Analysis

et al. 1999

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Vibrio parahaemolyticus is one of the most important food-borne pathogens in Taiwan, Japan, and other countries with long coastlines. This paper reports on the development of a new random amplified polymorphic DNA (RAPD) method for the molecular typing of this pathogen. The 10-mer primer 284 (5′-CAG GCG CAC A-3′) was selected to generate polymorphic amplification profiles of the genomic DNA at an annealing temperature of 38°C. A total of 308 clinical isolates of V. parahaemolyticus collected during food poisoning outbreaks in Taiwan, mostly occurring between 1993 and 1995, plus 11 environmental and clinical reference strains were analyzed by this RAPD method. A total of 41 polymorphic RAPD patterns were recognized, and these patterns were arbitrarily grouped into 16 types (A to P). Types A, B, C, D, and E were the major types, and subtypes C3, C5, E1, B1, D2, and A2 were the major patterns. The major types were phylogenetically more closely related to each other than to any of the minor types.

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Section: Resultsmentioning

confidence: 70%

Molecular Typing of Vibrio parahaemolyticus Isolates, Obtained from Patients Involved in Food Poisoning Outbreaks in Taiwan, by Random Amplified Polymorphic DNA Analysis

et al. 1999

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“…DNA was extracted from bacterial cells by a freeze/thaw method (Grossman and Ron 1975). PCR amplification was carried out according to the method described by Makino et al. (1994).…”

Section: Methodsmentioning

confidence: 99%

16S rRNA gene sequence-based analysis of clostridia related to conversion of germfree mice to the normal state

Momose

Maruyama

Iwasaki

et al. 2009

Journal of Applied Microbiology

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Aims: To determine phylogenetic groups of clostridia inhabiting the mouse intestine that are essential for normalization of germfree (GF) mice. Methods and Results: Using both the culture method and cloning, clostridia inhabiting the mouse intestine were isolated, and phylogenetic analysis based on 16S rRNA gene sequences was carried out. As a result, the isolates were found to have novel sequences, and no isolate was determined to be identical to previously known identified clostridia. Although the taxonomy of mouse intestinal clostridia was complex, many of them belonged to Clostridium clusters XIVa and IV in conventional (CV) and limited flora mice and ex‐germfree mice administered chloroform‐treated CV mouse faeces. The clostridia that belonged to cluster XIVa were most often present and showed the highest diversity. Conclusions: Clostridia belonging clusters XIVa and IV are dominant in the mouse intestine as in other gut ecosystems. The novel groups in these clusters are essential for normalization of GF mice. Significance and Impact of the Study: The results of this study can be applied in the strict control of mouse intestinal microbiota and will provide important information for normalization of GF mice and also for research on microbiology of the mouse intestine.

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“…Briefly amplified polymorphic DNA (RAPD) (15), pulsed field gel electrophoresis analysis (PFGE) and Southern transfer hybridization were performed. As arbitrary primers for RAPD analysis, A07 (5 '-TGCCTGCACCA-3 ') and A09 (5 '-CCGCAGTTAGAT-3 ') were used.…”

Section: Methodsmentioning

confidence: 99%

Detection and Genetical Characterization of Shiga Toxin‐Producing Escherichia coli from Wild Deer

Asakura

Makino

Shirahata

et al. 1998

Microbiology and Immunology

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Shiga toxin (Stx)-producing Escherichia coli (STEC) strains isolated from wild deer in Japan were examined. A total of 43 fecal samples were collected 4 times from 4 different sites around Obihiro City, Hokkaido, Japan, in June and July 1997. Seven STEC strains were isolated by PCR screening, all of them were confirmed by ELISA and Vero cell cytotoxicity assay to be producing only active Stx type 2 (Stx2). Moreover, they seemed to carry the hemolysin and eaeA genes of STEC 0157:H7, and some isolates harbored large plasmids which were similar to the 90-kilobase virulence plasmid of STEC 0157:H7. Based on their plasmid profiles, antibiotic resistance patterns, PCR-based DNA fingerprinting data obtained by using random amplified polymorphic DNA (RAPD) and the stx2 gene sequences, all isolates were divergent from each other except for 3 isolates from the first and second samplings. A DNA sequence analysis of representative isolates revealed that deer originating STEC strains were closely related to each other, but not to the Stx2-producing STEC strains isolated from a mass outbreak in Obihiro at the same time. A phylogenic analysis of the deduced Stx2 amino acid sequences demonstrated that three distinct clusters existed in the deer originating STEC strains and that the Stx of deer originating STEC was closely associated with that originating from humans, but not those of STEC originating from other animals. These results suggest that STEC contamination of deer carcasses should be considered as a potential source of human infection and adequate sanitary inspection of meat for human consumption is also essential for wild animals.

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PCR-based random amplified polymorphic DNA fingerprinting of Yersinia pseudotuberculosis and its practical applications

Cited by 54 publications

References 21 publications

Molecular Typing of Vibrio parahaemolyticus Isolates, Obtained from Patients Involved in Food Poisoning Outbreaks in Taiwan, by Random Amplified Polymorphic DNA Analysis

Molecular Typing of Vibrio parahaemolyticus Isolates, Obtained from Patients Involved in Food Poisoning Outbreaks in Taiwan, by Random Amplified Polymorphic DNA Analysis

16S rRNA gene sequence-based analysis of clostridia related to conversion of germfree mice to the normal state

Detection and Genetical Characterization of Shiga Toxin‐Producing Escherichia coli from Wild Deer

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