Amplification of the 16S-23S r DNA spacer region for rapid detection of Clostridium chauvoei and Clostridium septicum

Sasaki, Yohei; Yamamoto, Kazuhiro; Amimoto, Katsuhiko; Kojima, Akira; Ogikubo, Yasuaki; Norimatsu, Mari; Ogata, Hiroyuki; Tsutsumi, Yutaka

doi:10.1053/rvsc.2001.0495

Cited by 21 publications

(19 citation statements)

References 11 publications

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“…Parallel testing of the DNA isolated from the sample material with three different conventional PCR systems targeting the 16S-23S rRNA gene spacer region and the flagellin gene (13,14,16) yielded the same results as those obtained with the newly designed real-time PCR assay (data not shown). Thus, these discrepancies cannot be assigned to misidentification of the Clostridium species at the DNA level.…”

Section: Discussionmentioning

confidence: 54%

“…An IAC permits recognition of false-negative results due to inhibitory effects of the sample matrix (5). Conventional PCR assays for the detection of C. chauvoei published thus far do not include an IAC (1,6,8,13,14,15,16,18).…”

Section: Discussionmentioning

confidence: 99%

“…The rRNA database, which does not contain information about C. chauvoei and C. septicum, yielded an average of 9.43 operons for the clostridial species listed there (10). With respect to contamination levels of 10 7 to 10 5 CFU/g found in diseased muscle tissue (6,14), the detection limit of the proposed assay achieved in artificially contaminated muscle tissue (1.79 ϫ 10 3 CFU/g) suggested reliable detection of the pathogen in clinical specimens.…”

Section: Discussionmentioning

confidence: 99%

“…Conventional PCR systems target the 16S rRNA gene, the 16S-23S rRNA gene spacer region, or the flagellin gene (1,6,8,13,14,15,16,18). On one hand, conventional PCR bears the risk of yielding false-positive results due to contamination during post-PCR handling of the amplicons for detection.…”

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confidence: 99%

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Novel Real-Time PCR Assay for Simultaneous Detection and Differentiation of Clostridium chauvoei and Clostridium septicum in Clostridial Myonecrosis

et al. 2010

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A real-time PCR assay based on the 16S rRNA gene sequence was designed for differentiation of blacklegcausing Clostridium chauvoei and Clostridium septicum, a phylogenetically closely related bacterium responsible for malignant edema. In order to exclude false-negative results, an internal amplification control was included in the assay. A set of three probes, one specific for C. chauvoei, one specific for C. septicum, and one specific for both species, permitted unequivocal detection of C. chauvoei in tests of 32 Clostridium sp. strains and 10 non-Clostridium strains. The assay proved to be sensitive, detecting one genome of C. chauvoei or C. septicum per PCR and 1.79 ؋ 10 3 C. chauvoei cells/g artificially contaminated muscle tissue. In tests of 11 clinical specimens, the real-time PCR assay yielded the same results as an established conventional PCR method.

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Section: Discussionmentioning

confidence: 54%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Novel Real-Time PCR Assay for Simultaneous Detection and Differentiation of Clostridium chauvoei and Clostridium septicum in Clostridial Myonecrosis

et al. 2010

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“…C. septicum and C. chauvoei are genetically very similar, making them difficult to distinguish in biochemical testing. In the present study, however, the human fulminant gas gangrene case was confirmed to be caused by C. chauvoei after precise genetic characterizations including analyses of the 16S rRNA gene (6) and the 16S-23S rRNA gene intergenic spacer region (9,10). Therefore, strains from human patients with devastating clostridial infections that have been identified as C. septicum are worth checking out by further genetic identification, as was done in the present study.…”

Section: Case Reportmentioning

confidence: 47%

Human Fulminant Gas Gangrene Caused by Clostridium chauvoei

et al. 2008

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Advances in DNA Microarray Technology for the Detection of Foodborne Pathogens

Cremonesi¹,

Consolandi

Bellis

et al. 2010

Food Bioprocess Technol

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Hundreds of foodborne infection cases occur around the world, and up to one third of the population in industrialized nations suffers from foodborne illness each year. The advent of genetic-based technologies made feasible developing sensitive and specific screening tests for the detection of microbial pathogens. Microarray-based technologies represent an advance in nucleic acid testing methods whose main features include miniaturization, ability to parallelize sample processing, and ease of automation. Many applications, based on both commercial and custom arrays, have already been reported. The constant attempt to obtain reliable, sensitive, and robust methods has also driven the development of different molecular methods, relying on hybridization or on enzymatic techniques (such as minisequencing/extension or ligation). At the same time, probe design strategies and data analysis pipelines have been refined as well, in order to make result evaluations faster and less prone to errors. The high number of genetic information already available allowed reaching a resolution below the species level, being able to discriminate among serovars or strains, thanks to the careful choice of variable regions. Currently, the sensitivity of the assays can be as low as 1 CFU/ g for some bacteria after enrichment. In this review, we present an overview of the most important advances in microarray technologies in the foodborne pathogen detection field.

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Amplification of the 16S-23S r DNA spacer region for rapid detection of Clostridium chauvoei and Clostridium septicum

Cited by 21 publications

References 11 publications

Novel Real-Time PCR Assay for Simultaneous Detection and Differentiation of Clostridium chauvoei and Clostridium septicum in Clostridial Myonecrosis

Novel Real-Time PCR Assay for Simultaneous Detection and Differentiation of Clostridium chauvoei and Clostridium septicum in Clostridial Myonecrosis

Human Fulminant Gas Gangrene Caused by Clostridium chauvoei

Advances in DNA Microarray Technology for the Detection of Foodborne Pathogens

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