Propidium monoazide combined with real-time PCR for selective detection of viable Staphylococcus aureus in milk powder and meat products

From a public health point of view meat products contain high pathogenic risk factors. This is because they can be contaminated with Salmonella spp. and Staphylococcus aureus microorganisms, and, more importantly, antibiotic resistance has been reported in these microorganisms at an increasing frequency. To examine the presence of Salmonella spp. and Staphylococcus aureus in samples of raw and semi-cooked (chicken doner, meat doner, chicken, beef, and lamb products) meat from markets of Izmir and Balikesir, Turkey were analysed. The presence of microorganisms in the samples was determined by Real-Time PCR method using Salmonella spp. and S. aureus specific primers. Following Real-Time PCR, microorganisms were isolated by selective culture methods and biochemical tests from the positive meat samples and tested for their antibiotic susceptibility using the Kirby-Bauer Disk Diffusion Method. The antibiotic disc diffusion method showed that S. aureus was resistant to penicillin G, oxytetracycline, sulfamethoxazole, tetracycline, erythromycin, and ampicillin, whereas Salmonella spp. was resistant to penicillin G, sulfamethoxazole, erythromycin, and ampicillin. As these products are consumed frequently, their contamination with S. aureus (≥ 5 × 103 cfu/g) and Salmonella spp. can be a risk factor for food poisoning. The contamination of meat products’ with S. aureus and Salmonella spp. can be a risk factor for public health and the antibiotics to be preferred in illness treatment are of critical importance.

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Selective detection of viableEnterococcus faecalisusing propidium monoazide in combination with real-time PCR

The Journal of Korean Academy of Conservative Dentistry ◽

10.5395/jkacd.2008.33.6.537 ◽

2008 ◽

Vol 33 (6) ◽

pp. 537 ◽

Cited By ~ 2

Author(s):

Sinyoung Kim ◽

Seungjong Lee ◽

Euiseong Kim ◽

Deoggyu Seo ◽

Yoonjung Song ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Selective Detection ◽

Propidium Monoazide

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Quantitative Analysis of Staphylococcus aureus in Skimmed Milk Powder by Real-Time PCR

Journal of Veterinary Medical Science ◽

10.1292/jvms.67.1037 ◽

2005 ◽

Vol 67 (10) ◽

pp. 1037-1041 ◽

Cited By ~ 13

Author(s):

Tetsuya IKEDA ◽

Naoto TAMATE ◽

Keiji YAMAGUCHI ◽

Sou-ichi MAKINO

Keyword(s):

Staphylococcus Aureus ◽

Quantitative Analysis ◽

Real Time ◽

Real Time Pcr ◽

Milk Powder ◽

Skimmed Milk ◽

Skimmed Milk Powder

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Multiplex real-time PCR coupled with sodium dodecyl sulphate and propidium monoazide for the simultaneous detection of viable Listeria monocytogenes, Cronobacter sakazakii, Staphylococcus aureus and Salmonella spp. in milk

International Dairy Journal ◽

10.1016/j.idairyj.2020.104739 ◽

2020 ◽

Vol 108 ◽

pp. 104739 ◽

Cited By ~ 3

Author(s):

Hong Qin ◽

Xiuquan Shi ◽

Liang Yu ◽

Ke Li ◽

Jianwu Wang ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Listeria Monocytogenes ◽

Real Time ◽

Sodium Dodecyl Sulphate ◽

Real Time Pcr ◽

Sodium Dodecyl ◽

Simultaneous Detection ◽

Cronobacter Sakazakii ◽

Propidium Monoazide ◽

Salmonella Spp

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Selective detection of viable seed-borne Acidovorax citrulli by real-time PCR with propidium monoazide

Scientific Reports ◽

10.1038/srep35457 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 14

Author(s):

Qian Tian ◽

Jian-jun Feng ◽

Jie Hu ◽

Wen-jun Zhao

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Selective Detection ◽

Viable Seed ◽

Propidium Monoazide ◽

Acidovorax Citrulli

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Regulation of virulence and β-lactamase gene expression in Staphylococcus aureus isolates: cooperation of two-component systems in bloodstream superbugs

BMC Microbiology ◽

10.1186/s12866-021-02257-4 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Sanaz Dehbashi ◽

Hamed Tahmasebi ◽

Behrouz Zeyni ◽

Mohammad Reza Arabestani

Keyword(s):

Staphylococcus Aureus ◽

Real Time ◽

Real Time Pcr ◽

Quantitative Real Time Pcr ◽

Gene Expressions ◽

Resistance Development ◽

Component Systems ◽

Two Component ◽

Two Component Systems ◽

Lactamase Gene

Abstract Background Methicillin-resistant Staphylococcus aureus (MRSA)-bloodstream infections (BSI) are predominantly seen in the hospital or healthcare-associated host. Nevertheless, the interactions of virulence factor (VFs) regulators and β-lactam resistance in MRSA-BSI are unclear. This study aims to characterize the molecular relationship of two-component systems of VFs and the expression of the β-lactamase gene in MRSA-BSI isolates. In this study, 639 samples were collected from BSI and identified by phenotypic methods. We performed extensive molecular characterization, including SCCmec type, agr type, VFs gene profiles determinations, and MLST on isolates. Also, a quantitative real-time PCR (q-RT PCR) assay was developed for identifying the gene expressions. Results Ninety-one (91) S. aureus and 61 MRSA (67.0%) strains were detected in BSI samples. The presence of VFs and SCCmec genes in MRSA isolates were as follows: tst (31.4%), etA (18.0%), etB (8.19%), lukS-PVL (31.4%), lukF-PV (18.0%), lukE-lukD (16.3%), edin (3.2%), hla (16.3%), hlb (18.0%), hld (14.7%), hlg (22.9%), SCCmecI (16.3%), SCCmecII (22.9%), SCCmecIII (36.0%), SCCmecIV (21.3%), and SCCmecV (16.3%). Quantitative real-time PCR showed overexpression of mecRI and mecI in the toxigenic isolates. Moreover, RNAIII and sarA genes were the highest expressions of MRSA strains. The multi-locus sequence typing data confirmed a high prevalence of CC5, CC8, and CC30. However, ST30, ST22, and ST5 were the most prevalent in the resistant and toxigenic strains. Conclusion We demonstrated that although regulation of β-lactamase gene expressions is a significant contributor to resistance development, two-component systems also influence antibiotic resistance development in MRSA-BSI isolates. This indicates that resistant strains might have pathogenic potential. We also confirmed that some MLST types are more successful colonizers with a potential for MRSA-BSI.

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