Sequencing of an internal transcribed spacer region of 16S–23S rRNA gene and designing of PCR primers for the detection of Salmonella spp. in food

Chiu, Tasi-Hsin; Chen, Tong-Rong; Hwang, Wen-Zhe; Tsen, Hau‐Yang

doi:10.1016/j.ijfoodmicro.2004.04.005

Cited by 49 publications

(27 citation statements)

References 22 publications

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“…Ribosomal RNA genes evolve slowly and are highly conserved but it has been considered that the nucleotide sequences of the rRNA intergenic spacer regions are not as essential as the rRNA genes themselves and consequently not as highly conserved [20]. These properties made them a suitable site for developing specific PCR primers that can differentiate similar bacteria.…”

Section: Discussionmentioning

confidence: 99%

Detection of Pectobacteria Causal Agents of Potato Soft Rot in North Western Provinces of Iran

Azadmanesh¹

2012

J Plant Pathol Microb

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Various bacteria belonging to Enterobacteriaceae are known to be agents causing blackleg and soft rot in potato. In this study, a rapid and specific detection method was used to investigate bacteria in some northwestern provinces of Iran (Zanjan, Kordestan and eastern Azarbaijan). 26 strains were selected for the study that had been identified in previous studies as representative pathogens, they were as follows: 14 Pectobactrium carotovorum subsp carotovorum, 7 Dickeya chrysanthemi and 5 Pectobactrium atrosepticum. Primers Y1/Y2, Expccf/r, ADE1/2 and Eca1f/r, published as specific primers for the pathogens. They were tested to determine if they could be used for specific amplification of DNA in the strains. The expected specific fragment was not amplified in many strains or several non-specific bands, some close to those expected fragments were amplified. Specific primers for amplification of the DNA of pathogens were designed by sequencing the ITS region from representative strains and a PCR test was developed. In the PCR test a 300 bp fragment was amplified from all strains collected from the provinces. In specificity tests, no PCR products were obtained from other bacteria belonging to other genera.

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Section: Discussionmentioning

confidence: 99%

Detection of Pectobacteria Causal Agents of Potato Soft Rot in North Western Provinces of Iran

Azadmanesh¹

2012

J Plant Pathol Microb

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“…PCR primers targeting all Salmonella species were used to amplify DNA from both dust and filter samples (17)…”

Section: Methodsmentioning

confidence: 99%

Microbial Contents of Vacuum Cleaner Bag Dust and Emitted Bioaerosols and Their Implications for Human Exposure Indoors

Veillette

Knibbs

Pelletier

et al. 2013

Appl Environ Microbiol

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Vacuum cleaners can release large concentrations of particles, both in their exhaust air and from resuspension of settled dust. However, the size, variability, and microbial diversity of these emissions are unknown, despite evidence to suggest they may contribute to allergic responses and infection transmission indoors. This study aimed to evaluate bioaerosol emission from various vacuum cleaners. We sampled the air in an experimental flow tunnel where vacuum cleaners were run, and their airborne emissions were sampled with closed-face cassettes. Dust samples were also collected from the dust bag. Total bacteria, total archaea, Penicillium/Aspergillus, and total Clostridium cluster 1 were quantified with specific quantitative PCR protocols, and emission rates were calculated. Clostridium botulinum and antibiotic resistance genes were detected in each sample using endpoint PCR. Bacterial diversity was also analyzed using denaturing gradient gel electrophoresis (DGGE), image analysis, and band sequencing. We demonstrated that emission of bacteria and molds (Penicillium/Aspergillus) can reach values as high as 1E5 cell equivalents/min and that those emissions are not related to each other. The bag dust bacterial and mold content was also consistent across the vacuums we assessed, reaching up to 1E7 bacterial or mold cell equivalents/g. Antibiotic resistance genes were detected in several samples. No archaea or C. botulinum was detected in any air samples. Diversity analyses showed that most bacteria are from human sources, in keeping with other recent results. These results highlight the potential capability of vacuum cleaners to disseminate appreciable quantities of molds and human-associated bacteria indoors and their role as a source of exposure to bioaerosols.

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“…To our knowledge, so far functional gene diversity has not been studied in food-associated matrices. However, several applications of gene functional analysis have been reported for detection of and simultaneous discrimination among food-borne human pathogens (Chizhikov et al, 2001;Volokhov et al, 2002;Chiu et al, 2005).…”

Section: Functional Genesmentioning

confidence: 99%

Recent advances in molecular techniques to study microbial communities in food-associated matrices and processes

2008

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Sequencing of an internal transcribed spacer region of 16S–23S rRNA gene and designing of PCR primers for the detection of Salmonella spp. in food

Cited by 49 publications

References 22 publications

Detection of Pectobacteria Causal Agents of Potato Soft Rot in North Western Provinces of Iran

Detection of Pectobacteria Causal Agents of Potato Soft Rot in North Western Provinces of Iran

Microbial Contents of Vacuum Cleaner Bag Dust and Emitted Bioaerosols and Their Implications for Human Exposure Indoors

Recent advances in molecular techniques to study microbial communities in food-associated matrices and processes

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