Detection of the Salmonella typhi bacteria in contaminated egg using real-time PCR to develop rapid detection of food poisoning bacteria

Nurjayadi, Muktiningsih; Pertiwi, Yuniariana; Islami, N.; Azizah, N.; Efrianti, Ulfi Rahma; Saamia, V.; Wiranatha, I. M.; Nastassya, L.; El-Enshasye, H. Ali

doi:10.1016/j.bcab.2019.101214

Cited by 23 publications

(13 citation statements)

References 9 publications

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“…To circumvent the limitations of conventional serotyping, many studies have focused on developing real-time PCR with multiple reactions or multiplex real-time PCR because of their ease of use (24,25), coupling with other methods based on Luminex, CRISPR (26), or WGS. The drawbacks of these methods include either the need of bioinformatics analysis and/or gel electrophoresis after PCR ampli cation, or reliance on special and costly instruments, or procedures being time consuming or complex, which limit their widespread use in clinical settings.…”

Section: Discussionmentioning

confidence: 99%

Rapid and Specific Differentiation of Salmonella enterica serotypes Typhi and Paratyphi by Multicolor Melting Curve Analysis

Jiang,

Cai

et al. 2024

Preprint

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Rapid and accurate identification of Salmonella enterica serotypes Typhi and Paratyphi (A, B and C), the causal agents of enteric fever, is critical for timely treatment, case management and evaluation of health policies in low and middle-income countries where the disease still remains a serious public health problem. The present study describes the development of a multiplex assay (EFMAtyping) for simultaneous identification of pathogens causing typhoid and paratyphoid fever in a single reaction by the MeltArray approach, which could be finished within 2.5 h. Seven specific genes were chosen for differentiation of typhoidal and nontyphoidal Salmonella. All gene targets were able to be detected by the EFMAtyping assay, with expected Tm values and without cross-reactivity to other relevant Salmonella serovars. The limit of detection (LOD) for all gene targets was 50 copies per reaction. The LOD reached 102-103 CFU/ml for each pathogen in simulated clinical samples. The largest standard deviation value for mean Tm was below 0.5°C. This newly developed EFMAtyping assay was further evaluated by testing 551 clinical Salmonella isolates, corroborated in parallel by the traditional Salmonella identification workflow, and serotype prediction was enabled by whole-genome sequencing. Compared to the traditional method, our results exhibited 100% of specificity and greater than 96% of sensitivity with a kappa correlation ranging from 0.96 to 1.00. Thus, the EFMAtyping assay provides a rapid, high throughput, and promising tool for public health laboratories to monitor typhoid and paratyphoid fever.

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Section: Discussionmentioning

confidence: 99%

Rapid and Specific Differentiation of Salmonella enterica serotypes Typhi and Paratyphi by Multicolor Melting Curve Analysis

Jiang,

Cai

et al. 2024

Preprint

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“…The quantitative real-time PCR assay for ure C gene was used to calculate the H. pylori number to determine the limit of detection of the developed RUT. The PCR product obtained from the real-time PCR reaction of each bacterial species has an appropriate melting temperature (Tm) that was displayed with DNA peak in the graph, which determines the conditions under which these will bind to target gene [ 21 ]. The results of the melting point curve show that there was no mispriming in the test; the use of Tm at 78°C ensures that the primer used only amplifies the ure C target gene, as indicated by the formation of a single peak ( Figure-3d ).…”

Section: Discussionmentioning

confidence: 99%

Development of an easy-to-use urease kit for detecting Helicobacter pylori in canine gastric mucosa

et al. 2021

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Background and Aim: Helicobacter pylori is an important pathogen in humans and animals involved in chronic gastritis, leading to the development of gastric cancer. Urease produced by H. pylori is an enzyme that promotes bacterial colonization and can be used clinically as a biomarker of H. pylori infection as part of a rapid urease test (RUT). A test with high specificity (95-100%) would be more convenient and faster than histopathology, bacterial culture, and polymerase chain reaction (PCR). The aim of this study was to develop a simple, cheap, and fast kit for detecting H. pylori infection in the gastric mucosa of canines, which can be used in clinical practice for diagnosing infection with this bacterium. Materials and Methods: The RUT assays developed were prepared using 1% agar, 1% sodium phosphate monobasic, and 1% urea followed by the addition of 3% methyl red indicator. The cutoff value of sensitivity of the RUT assay was established using the urease of H. pylori ATCC 43504 and color change was monitored for 24 h. Comparisons of the sensitivity to H. pylori ATCC 43504 were made between the developed RUT assays and the Hp Fast™ commercial kit. Then, the limit of detection for H. pylori ATCC 43504 number was analyzed by the SYBR Green real-time PCR assay to measure the copy number of the ureC gene. Gastric biopsy samples from the antrum, body, and fundus of the stomach were collected from eight canines presenting with vomiting and gastroenteritis. Analyses were performed on fresh samples using the developed RUT assays and the Hp Fast™ commercial kit, which were read within 24 h; then, the results were confirmed with SYBR Green real-time PCR. The specificity of the RUT assays was tested with a number of different bacteria, including Staphylococcus pseudintermedius, Proteus spp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterococcus spp., Escherichia coli, and Salmonella spp.; H. pylori ATCC 43504 was used as a positive control. Results: The results showed that the developed assays were sensitive to the urease enzyme at 0.1 mg/mL. The lowest detection limit of this assay for H. pylori ATCC 43504 was found to be 102 copies at 30 min. The sensitivity of detection of H. pylori in gastric biopsies of canines occurred in a minimum of 30 min. The RUT showed similar results to the Hp Fast™ commercial kit. In the developed RUT, the color change of the test from red to yellow could be clearly distinguished between the color of the positive test and the negative one; however, in the commercial Hp Fast™, it was difficult to observe the gel color change in the negative pH range of 5.8 and the positive pH of 6.5. The developed RUT was specific for H. pylori and did not detect any of the other tested bacteria. The test kit can also be stored for 6 months at 4°C. Conclusion: The sensitivity of the developed assays allowed the detection of urease enzyme at a minimum concentration of 0.1 mg/mL. Our RUT could also detect H. pylori from one in eight canine specimens at a minimum of 102 copies within 30 min. This RUT is specific to H. pylori as it did not detect any of the other tested bacteria.

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“…In the EU, nearly half of all cases of diarrhea due to contamination by Shiga toxin-producing Escherichia coli require hospitalization [ 67 ]. Food poisoning in Salmonella causes salmonellosis [ 68 ]. S. aureus staphylococcal enterotoxins frequently cause staphylococcal food poisoning during food contamination.…”

Section: Food Spoilagementioning

confidence: 99%

Potential of Syzygnium polyanthum as Natural Food Preservative: A Review

Julizan

Ishmayana

Zainuddin

et al. 2023

Foods

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Food preservation is one of the strategies taken to maintain the level of public health. Oxidation activity and microbial contamination are the primary causes of food spoilage. For health reasons, people prefer natural preservatives over synthetic ones. Syzygnium polyanthum is widely spread throughout Asia and is utilized as a spice by the community. S. polyanthum has been found to be rich in phenols, hydroquinones, tannins, and flavonoids, which are potential antioxidants and antimicrobial agents. Consequently, S. polyanthum presents a tremendous opportunity as a natural preservative. This paper reviews recent articles about S. polyanthum dating back to the year 2000. This review summarizes the findings of natural compounds presented in S. polyanthum and their functional properties as antioxidants, antimicrobial agents, and natural preservatives in various types of food.

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Detection of the Salmonella typhi bacteria in contaminated egg using real-time PCR to develop rapid detection of food poisoning bacteria

Cited by 23 publications

References 9 publications

Rapid and Specific Differentiation of Salmonella enterica serotypes Typhi and Paratyphi by Multicolor Melting Curve Analysis

Rapid and Specific Differentiation of Salmonella enterica serotypes Typhi and Paratyphi by Multicolor Melting Curve Analysis

Development of an easy-to-use urease kit for detecting Helicobacter pylori in canine gastric mucosa

Potential of Syzygnium polyanthum as Natural Food Preservative: A Review

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