A duplex real-time NASBA assay targeting serotype-specific gene for rapid detection of viable S. enterica serovar Paratyphi C in retail foods of animal origin

2022 ◽  
Author(s):  
Zhai Ligong ◽  
Liu Hongxia ◽  
Li Junjie ◽  
Zhaoxin Lu ◽  
Xiaomei Bie
Keyword(s):  
Real Time ◽  
Molecular Beacon ◽  
Detection Method ◽  
Simultaneous Detection ◽  
Animal Origin ◽  
Specific Gene ◽  
Salmonella Spp ◽  
Viable Cells ◽  
Food Of Animal Origin ◽  
Nasba Assay

Salmonella enterica serovars Paratyphi C is highly adapted to humans and can cause a typhoid-like disease with high mortality rates. In this study, three serovar-specific genes were determined for S. Paratyphi C, SPC_0871，SPC_0872, and SPC_0908, by comparative genomics method. Based on SPC_0908 and xcd gene for testing Salmonella spp., we have developed a duplex real-time nucleic acid sequence-based amplification (real-time NASBA) with molecular beacon approach for simultaneous detection of viable cells of Salmonella spp. and serotype Paratyphi C. The test selectively and consistently detected 53 Salmonella spp. (representing 31 serotypes) and 18 non-Salmonella strains. Additionally, the method showed high resistance to interference by natural background flora in pork and chicken samples. The sensitivity of the established approach was determined to be 4.89 CFU/25 g in artificially contaminated pork and chicken samples after pre-enrichment. We propose this NASBA-based protocol as a potential detection method for Salmonella spp. and serotype Paratyphi C in food of animal origin.

scholarly journals Citrobacter braakii Yield False-Positive Identification as Salmonella, a Note of Caution

Foods ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 2177
Author(s):  
Joanna Pławińska-Czarnak ◽  
Karolina Wódz ◽  
Magdalena Kizerwetter-Świda ◽  
Tomasz Nowak ◽  
Janusz Bogdan ◽  
...  
Keyword(s):  
False Positive ◽  
Salmonella Enterica ◽  
Meat Products ◽  
Animal Origin ◽  
Biochemical Tests ◽  
Salmonella Spp ◽  
Isolation And Identification ◽  
False Positive Identification ◽  
Food Of Animal Origin ◽  
Meat Samples

Background: Globally, Salmonella enterica is one of the leading causes of foodborne illness in humans. Food of animal origin is obligatorily tested for the presence of this pathogen. Unfortunately, in meat and meat products, this is often hampered by the presence of background microbiota, which may present as false-positive Salmonella. Methods: For the identification of Salmonella spp. from meat samples of beef, pork, and poultry, the authorized detection method is PN-EN ISO 6579-1:2017-04 with the White–Kauffmann–Le Minor scheme, two biochemical tests: API 20E and VITEK II, and a real-time PCR-based technique. Results: Out of 42 presumptive strains of Salmonella, 83.3% Salmonella enterica spp. enterica, 14.3% Citrobacter braakii, and 12.4% Proteus mirabilis were detected from 180 meat samples. Conclusions: Presumptive strains of Salmonella should be identified based on genotypic properties such as DNA-based methods. The aim of this study was the isolation and identification of Salmonella spp. from miscellaneous meat sorts: beef, pork, and poultry.

scholarly journals Real-Time Nucleic Acid Sequence-Based Amplification Assay for Detection of Hepatitis A Virus

2005 ◽  
Vol 71 (11) ◽  
pp. 7113-7116 ◽  
Author(s):  
Khaled H. Abd El Galil ◽  
M. A. El Sokkary ◽  
S. M. Kheira ◽  
Andre M. Salazar ◽  
Marylynn V. Yates ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
Hepatitis A ◽  
Molecular Beacon ◽  
Hepatitis A Virus ◽  
Immunomagnetic Separation ◽  
Nucleic Acid Sequence ◽  
The Real ◽  
Nasba Assay ◽  
Environmental Pathogens

ABSTRACT A nucleic acid sequence-based amplification (NASBA) assay in combination with a molecular beacon was developed for the real-time detection and quantification of hepatitis A virus (HAV). A 202-bp, highly conserved 5′ noncoding region of HAV was targeted. The sensitivity of the real-time NASBA assay was tested with 10-fold dilutions of viral RNA, and a detection limit of 1 PFU was obtained. The specificity of the assay was demonstrated by testing with other environmental pathogens and indicator microorganisms, with only HAV positively identified. When combined with immunomagnetic separation, the NASBA assay successfully detected as few as 10 PFU from seeded lake water samples. Due to its isothermal nature, its speed, and its similar sensitivity compared to the real-time RT-PCR assay, this newly reported real-time NASBA method will have broad applications for the rapid detection of HAV in contaminated food or water.

Next day Salmonella spp. detection method based on real-time PCR for meat, dairy and vegetable food products

2014 ◽  
Vol 184 ◽  
pp. 113-120 ◽  
Author(s):  
David Rodriguez-Lazaro ◽  
Patricia Gonzalez-García ◽  
Elisabetta Delibato ◽  
Dario De Medici ◽  
Rosa Maria García-Gimeno ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Detection Method ◽  
Food Products ◽  
Salmonella Spp ◽  
Vegetable Food

scholarly journals Epidemiological Study on Prevalence, Serovar Diversity, Multidrug Resistance, and CTX-M-Type Extended-Spectrum β-Lactamases of Salmonella spp. from Patients with Diarrhea, Food of Animal Origin, and Pets in Several Provinces of China

2020 ◽  
Vol 64 (7) ◽  
Author(s):  
Wei Wang ◽  
Li Zhao ◽  
Yujie Hu ◽  
Tania Dottorini ◽  
Séamus Fanning ◽  
...  
Keyword(s):  
Multidrug Resistant ◽  
Quinolone Resistance ◽  
Animal Origin ◽  
Salmonella Spp ◽  
Fecal Samples ◽  
Content Type ◽  
Extended Spectrum ◽  
Food Of Animal Origin ◽  
High Prevalence ◽  
Almost All

ABSTRACT A total of 2,283 Salmonella isolates were recovered from 18,334 samples, including samples from patients with diarrhea, food of animal origin, and pets, across 5 provinces of China. The highest prevalence of Salmonella spp. was detected in chicken meats (39.3%, 486/1,237). Fifteen serogroups and 66 serovars were identified, with Salmonella enterica serovars Typhimurium and Enteritidis being the most dominant. Most (85.5%, 1,952/2,283) isolates exhibited resistance to ≥1 antimicrobial, and 56.4% were multidrug resistant (MDR). A total of 222 isolates harbored extended-spectrum β-lactamases (ESBLs), and 200 of these were of the CTX-M type and were mostly detected in isolates from chicken meat and turtle fecal samples. Overall, eight blaCTX-M genes were identified, with blaCTX-M-65, blaCTX-M-123, blaCTX-M-14, blaCTX-M-79, and blaCTX-M-130 being the most prevalent. In total, 166 of the 222 ESBL-producing isolates had amino acid substitutions in GyrA (S83Y, S83F, D87G, D87N, and D87Y) and ParC (S80I), while the plasmid-mediated quinolone resistance (PMQR)-encoding genes oqxA, oqxB, qepA, qnrB, and qnrS were detected in almost all isolates. Of the 15 sequence types (STs) identified in the 222 ESBLs, ST17, ST11, ST34, and ST26 ranked among the top 5 in number of isolates. Our study revealed considerable serovar diversity and a high prevalence of the co-occurrence of MDR determinants, including CTX-M-type ESBLs, quinolone resistance-determining region (QRDR) mutations, and PMQR genes. This is the first report of CTX-M-130 Salmonella spp. from patients with diarrhea and QRDR mutations from turtle fecal samples. Our study emphasizes the importance of actions, both in health care settings and in the veterinary medicine sector, to control the dissemination of MDR, especially the CTX-M-type ESBL-harboring Salmonella isolates.

Monitoring Italian establishments exporting food of animal origin to third countries: SSOP compliance and Listeria monocytogenes and Salmonella spp. contamination

Food Control ◽  
2021 ◽  
Vol 121 ◽  
pp. 107584
Author(s):  
Salvatore Antoci ◽  
Luigi Iannetti ◽  
Gabriella Centorotola ◽  
Vicdalia Aniela Acciari ◽  
Francesco Pomilio ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Animal Origin ◽  
Salmonella Spp ◽  
Food Of Animal Origin

scholarly journals Methods of Detecting Meat Species in Food of Animal Origin

2018 ◽  
Vol 75 (2) ◽  
pp. 125
Author(s):  
Lavinia-Maria CHIŞ ◽  
Dan-Cristian VODNAR
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Meat Product ◽  
Meat Products ◽  
Ease Of Use ◽  
Animal Origin ◽  
Processed Meat ◽  
Elisa Method ◽  
Food Of Animal Origin ◽  
Pcr Method

An important factor in the detection of falsification is the control of the composition of the meat at each stage of manufacturing the product. The PCR method is based on the study of proteins and meat nucleic acids used in food for the detection of animal species. Another technique is the Elisa method that works on the principle of identification and measurement of the quantity of molecules in a sample. There are several types of Elisa to increase specificity due to differences in structure and sample characteristics. By comparing the two methods used to identify the processed meat product species, Real Time PCR had the highest prediction as results. However, the Elisa method is more time efficient and easier to use. Real Time PCR is effective in identifying processed meat products that require low detection. The Elisa Kit is useful because of the ease of use.

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