Development of a rapid detection system for opportunistic pathogenic Cronobacter spp. in powdered milk products

Zimmermann, Jennifer; Schmidt, Herbert; Loessner, Martin J.; Weiß, Agnes

doi:10.1016/j.fm.2014.02.010

Cited by 31 publications

(30 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The virulence of these pathogens differs, and C. sakazakii causes the majority of serious neonatal and infant infections (Jackson et al, 2014;Zimmermann et al, 2014). Very few reports have described PCR assays for detection of C. sakazakii; thus, it is critical to develop methods targeting this important species of Cronobacter.…”

Section: Introductionmentioning

confidence: 99%

Mining for sensitive and reliable species-specific primers for PCR for detection of Cronobacter sakazakii by a bioinformatics approach

Chen

Tao

Bie

et al. 2015

Journal of Dairy Science

View full text Add to dashboard Cite

Although several studies have reported PCR assays for distinguishing Cronobacter sakazakii from other species in the genus, reports regarding assay sensitivity and specificity, as well as applications for food testing, are lacking. Hence, the objective of this study was to develop a sensitive and reliable PCR-based method for detection of C. sakazakii by screening for specific target genes. The genome sequence of C. sakazakii in the GenBank database was compared with that of other organisms using BLAST. Thirty-eight DNA fragments unique to C. sakazakii were identified, and primers targeting these sequences were designed. Finally, 3 primer sets (CS14, CS21, and CS38) were found to be specific for C. sakazakii by PCR verification. The detection limit of PCR assays using the 3 pairs of primers was 1.35 pg/μL, 135 fg/μL, and 135 fg/μL, respectively, for genomic DNA, and 5.5×10(5), 5.5×10(3), 5.5×10(3) cfu/mL, respectively, using pure cultures of the bacteria, compared with 13.5 pg/μLand 5.5×10(5) cfu/mLfor primer set SpeCronsaka, which has been previously described. Cronobacter sakazakii were detected in artificially contaminated powdered infant formula (PIF) by PCR using primer sets CS21 and CS38 after 8h of enrichment. The detection limit was 5.5×10(-1) cfu/10g of PIF. Thus, the PCR assay can be used for rapid and sensitive detection of C. sakazakii in PIF.

show abstract

Section: Introductionmentioning

confidence: 99%

Mining for sensitive and reliable species-specific primers for PCR for detection of Cronobacter sakazakii by a bioinformatics approach

Chen

Tao

Bie

et al. 2015

Journal of Dairy Science

View full text Add to dashboard Cite

show abstract

“…The results were consistent with a previous study conducted by Zimmermann et al . (), in which C. sakazakii DSM 4485 is detectable at 0.01 cfu/g.…”

Section: Discussionmentioning

confidence: 99%

“…(Zimmermann et al . ) As well as the six species with medical significance, Cronobacter condimenti has not been associated with human illness (Cruz‐Cordova et al . ).…”

Section: Introductionmentioning

confidence: 99%

Simultaneous detection and identification of pathogenic Cronobacter species by high‐resolution melting analysis in powdered infant formulas

et al. 2017

Int J of Dairy Tech

View full text Add to dashboard Cite

A combination method of real-time PCR and high-resolution melting (HRM) analysis for the rapid detection and specific classification of six pathogenic Cronobacter species based on gene cgcA was developed. HRM profiles with distinct T m (melting temperature) peaks were consistently obtained with each species, represented by a single peak ranging from 86.02 to 86.80°C. The detection limit ranged from 0.1 to 1 pg per reaction. Desiccated Cronobacter sakazakii and Cronobacter muytjensii of 2 cfu/25 g were successfully detected after 4-week storage at room temperature. The newly developed method provided a molecular tool for direct detection and simultaneous identification of pathogenic Cronobacter species in powdered infant formulae.

show abstract

“…According to the molecular identification, Cronobacteria classifies into seven species: Cronobacter sakazakii , Cronobacter muytjensii , Cronobacter malonaticus , Cronobacter universalis , Cronobacter condiment , Cronobacter dublinensis , & Cronobacter turicensis (Joseph, Hariri, & Forsythe, ). Among them, C. malonaticus , C. turicensis and C. sakazakii are very common species that have been separated from human infections (Jackson, Sonbol, Masood, & Forsythe, ; Zimmermann, Schmidt, Loessner, & Weiss, ).…”

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

View full text Add to dashboard Cite

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.

show abstract

Development of a rapid detection system for opportunistic pathogenic Cronobacter spp. in powdered milk products

Cited by 31 publications

References 42 publications

Mining for sensitive and reliable species-specific primers for PCR for detection of Cronobacter sakazakii by a bioinformatics approach

Mining for sensitive and reliable species-specific primers for PCR for detection of Cronobacter sakazakii by a bioinformatics approach

Simultaneous detection and identification of pathogenic Cronobacter species by high‐resolution melting analysis in powdered infant formulas

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

Contact Info

Product

Resources

About