Use of novel species-specific PCR primers targeted to DNA gyrase subunit B (gyrB) gene for species identification of the Cronobacter sakazakii and Cronobacter dublinensis

Huang, Chien‐Hsun; Chang, Mu-Tzu; Huang, Lina

doi:10.1016/j.mcp.2012.08.004

Cited by 23 publications

(20 citation statements)

References 27 publications

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“…The gyrB gene (encodes the subunit B protein of a type II DNA topoisomerase) has a crucial role in DNA replication, and is distributed universally among bacterial species. It has been suggested that this gene is an excellent molecular marker for bacterial species identification (Huang et al, ). The results of the present study showed that gyrB gene had high specificity in the identification of C. sakazakii .…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, it is essential to study other genes and primer for the detection of C. sakazakii by molecular methods. There are many previous studies on various genes of C. sakazakii such as wzx (Jarvis et al, ), rpoB (Stoop, Lehner, Iversen, Fanning, & Stephan, ), cgcA (Carter et al, ), gyrB (Huang, Chang, & Huang, ) , dnaG (Seo & Brackett, ), wehI , wehC (Mullane et al, ).…”

Section: Introductionmentioning

confidence: 99%

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A novel primer targeted gyrB gene for the identification of Cronobacter sakazakii in powdered infant formulas (PIF) and baby foods in Iran

Mashoufi

Ghazvini

Hashemi

et al. 2018

Journal of Food Safety

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The aim of this study was to perform a molecular identification and design a polymerase chain reaction (PCR) method based on a specific gene (gyrase subunit B [gyrB]) for rapid detection of Cronobacter sakazakii. In this study, from February 2017 to January 2018, 100 powdered infant formula milks (PIF1‐6) and 100 baby food items (BF1‐8) (total number = 200 samples) were purchased from different commercial brands from different pharmacies of the Mashhad city, Iran from different pharmacies of the Mashhad city, Iran. After isolation of Cronobacter, DNA extraction and PCR assay were performed to detect and confirm genus and species of isolated bacteria with 16S ribosomal RNA (16S rRNA) and designed gyrB primers, respectively. The abundance of C. sakazakii in PIFs and baby foods by culture method were 5/100 (5%) and 8/100 (8%), respectively; also, the final analysis based on gyrB primer pairs in PIF and baby food showed contamination rates of 0/100 (0%) and 3/100 (3%), respectively. These findings indicated that Iranian baby foods were a real threat for children's health. It is also recommended that this designed primer could be used for detection of Cronobacter sakazakii in these products. Practical application Cronobacter sakazakii is an opportunistic bacteria that is known worldwide as a foodborne pathogen especially in Infant Formula and baby foods, therefore the quality of these products has to be evaluated by a rapid, sensitive and specific method. Our findings indicated that Iranian baby foods were a threat for children's health so implementation of adequate sanitary measures to prevent Cronobacter sakazakii illness is necessary. It was also recommended that the designed primer based on gyr B gene could be used for specific detection of Cronobacter sakazakii in these products.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A novel primer targeted gyrB gene for the identification of Cronobacter sakazakii in powdered infant formulas (PIF) and baby foods in Iran

Mashoufi

Ghazvini

Hashemi

et al. 2018

Journal of Food Safety

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show abstract

“…GyrB was also found to be a very promising and useful alternative target for identification and taxonomic analysis for the Bacillus subtilis group that consists of eight very closely related species [97]. Similar approaches have been used to overcome limitations of rRNA-based diagnostic using a variety of additional targets, including but not limited to gyrB for various Cronobacter species [98], rpoB for Legionella [99] or even a combination of genes for some species including Acetinobacter spp. [100].…”

Section: Nonribosomal Nucleic Acid Marker Moleculesmentioning

confidence: 99%

Nucleic acid detection technologies and marker molecules in bacterial diagnostics

Scheler

Glynn

Kurg

2014

Expert Review of Molecular Diagnostics

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There is a growing need for quick and reliable methods for microorganism detection and identification worldwide. Although traditional culture-based technologies are trustworthy and accurate at a relatively low cost, they are also time- and labor-consuming and are limited to culturable bacteria. Those weaknesses have created a necessity for alternative technologies that are capable for faster and more precise bacterial identification from medical, food or environmental samples. The most common current approach is to analyze the nucleic acid component of analyte solution and determine the bacterial composition according to the specific nucleic acid profiles that are present. This review aims to give an up-to-date overview of different nucleic acid target sequences and respective analytical technologies.

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“…A pair of novel species‐specific PCR primers targeted to DNA gyrase subunit B ( gyrB ) gene for species identification of the C. sakazakii and C. dublinensis has been successfully used (Huang et al . ). It was found that different primer pairs based on the rpoB sequences of the six Cronobacter species type strains were designed (Stoop et al .…”

Section: Introductionmentioning

confidence: 97%

“…; Huang et al . ). Therefore, it is necessary to develop a method only for the detection of C. sakazakii strain in PIF.…”

Section: Introductionmentioning

confidence: 97%

Development of a PCR Assay for Rapid Detection of Cronobacter sakazakii from Powdered Infant Formula Using a Target Sequence Identified by Comparative Genomic Analysis

Chen

Ren

et al. 2015

Journal of Food Safety

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The occurrence of outbreaks of Cronobacter sakazakii causing necrotizing meningitis in China highlights the need for strain characterization of this pathogenic species. In this study, SMM system was utilized to mine for new molecular markers of C. sakazakii. One C. sakazakii‐specific CDS (>ref|NC_009778.1|:c1077329‐1076055) with a length of 1,275 bp was used to design a primer set. A PCR assay was developed and optimized to detect C. sakazakii. The PCR assay amplified a 498 bp DNA product from all 43 C. sakazakii strains tested but not from 30 other bacteria strains. The detection limit was 7.11 pg/PCR (equivalent 1,422 genomic copies) when the genomic DNA of C. sakazakii ATCC 29544 was 10‐fold diluted. Three powdered infant formulas from different commercial brands were inoculated with the strain ATCC 29544 at the level of 56 cells contamination, and the target fragments (498 bp) were produced after samples were enriched for 6 h at 37C.Thirty food samples were detected simultaneously by this PCR assay and the conventional method, and a coincident detecting result was found between the PCR assay and the conventional method. This method provides a useful tool for rapid identification of C. sakazakii in food and environmental matrices. Practical Applications The gold standard based on the FDA‐recommended methods for isolation and identification of Cronobacter sakazakii from powdered infant formula is time consuming and labor intensive. A PCR assay has been established to detect C. sakazakii. This method provides a useful tool for rapid and accurate identification of C. sakazakii in food when a large number of food samples need to be rapidly isolated and identified.

show abstract

Use of novel species-specific PCR primers targeted to DNA gyrase subunit B (gyrB) gene for species identification of the Cronobacter sakazakii and Cronobacter dublinensis

Cited by 23 publications

References 27 publications

A novel primer targeted gyrB gene for the identification of Cronobacter sakazakii in powdered infant formulas (PIF) and baby foods in Iran

A novel primer targeted gyrB gene for the identification of Cronobacter sakazakii in powdered infant formulas (PIF) and baby foods in Iran

Nucleic acid detection technologies and marker molecules in bacterial diagnostics

Development of a PCR Assay for Rapid Detection of Cronobacter sakazakii from Powdered Infant Formula Using a Target Sequence Identified by Comparative Genomic Analysis

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