Detection of viable Salmonella Typhi by reverse transcription-multiplex polymerase chain reaction

Phumkhachorn, Parichat; Rattanachaikunsopon, Pongsak

doi:10.9755/ejfa.2016-12-1867

Cited by 8 publications

(6 citation statements)

References 18 publications

(21 reference statements)

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“…Some of the recently published methods for the detection of S . typhi in blood, water samples, and food are, lateral flow immunoassay (LFIA) using antibody-coated gold nanoparticles [ 13 ], uniplex and, multiplex PCR [ 14 ], reverse transcriptase multiplex PCR (RT-MPCR) [ 15 ] and gel electrophoresis [ 16 ]. The limit of detection (LOD) of these above methods ranges between 500 to 10 4 CFU/mL.…”

Section: Introductionmentioning

confidence: 99%

Highly-sensitive detection of Salmonella typhi in clinical blood samples by magnetic nanoparticle-based enrichment and in-situ measurement of isothermal amplification of nucleic acids

et al. 2018

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Enteric fever continues to be a major cause of mortality and morbidity globally, particularly in poor resource settings. Lack of rapid diagnostic assays is a major driving factor for the empirical treatment of enteric fever. In this work, a rapid and sensitive method ‘Miod’ ‘has been developed. Miod includes a magnetic nanoparticle-based enrichment of target bacterial cells, followed by cell lysis and loop-mediated isothermal amplification (LAMP) of nucleic acids for signal augmentation along with concurrent measurement of signal via an in–situ optical detection system. To identify positive/negative enteric fever infections in clinical blood samples, the samples were processed using Miod at time = 0 hours and time = 4 hours post-incubation in blood culture media. Primers specific for the STY2879 gene were used to amplify the nucleic acids isolated from S. typhi cells. A limit of detection of 5 CFU/mL was achieved. No cross-reactivity of the primers were observed against 106 CFU/mL of common pathogenic bacterial species found in blood such as E. coli, P. aeruginosa, S. aureus, A. baumanni, E. faecalis, S. Paratyphi A and K. pneumonia. Miod was tested on 28 human clinical blood samples. The detection of both pre-and post-four-hours incubation confirmed the presence of viable S. typhi cells and allowed clinical correlation of infection. The positive and negative samples were successfully detected in less than 6 hours with 100% sensitivity and specificity.

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Section: Introductionmentioning

confidence: 99%

Highly-sensitive detection of Salmonella typhi in clinical blood samples by magnetic nanoparticle-based enrichment and in-situ measurement of isothermal amplification of nucleic acids

et al. 2018

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“…Previous research rarely used blood samples as the object to detect S. typhi (Nader et al, 2015;Shanmugasamy et al, 2011;Salehi et al, 2012). Phumkhachorn and Rattanachaikunsopon (2017) show that the Inv A gene has a specific sensitivity to detect S. typhi. While the Fim C gene can be used to detect salmonella from fecal samples (Jawad and Al-hamadani, 2011).…”

Section: Resultsmentioning

confidence: 89%

Detection of Salmonella typhi Using Multiplex and Monoplex PCR in Tifoid Fever Patients

Aliviameita

Agustin²,

Puspita³

et al. 2019

Med.Lab.Tech.J.

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Typhoid fever is a disease caused by Salmonella typhi infection. This disease is still a global problem. The purpose of this study was to develop a rapid detection method for Salmonella using molecular methods utilizing Inv A and Fim-C Genes. Two methods compared, which are Multiplex and monoplex PCR. The sample is in the form of the patient's blood, which is stored at 3 ml EDTA vacutainer tube consisting of 10 positive samples and five negative samples. The criteria for samples used previously have tested by widal/ immunoserology (titer more than 1/160). The results showed that multiplex PCR for detection of Salmonella using the Inv A gene and fim c gene not recommended because too many bands produced. The application of monoplex in the Inv A gene gives better results than the Fim C gene. In conclusion, the monoplex application on Inv A gene recommended than Fim C gene used to detect S. typhi in human blood samples.

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“…This is because; intracellular gene exchange between pathogens is possible when antimicrobial resistant Salmonella translocate to the mesenteric lymph nodes and other systemic sites from the gut via the intestine [25]. Consequently, the detection of the invA gene in all human isolates as reported by several studies may be possible through intracellular conjugative virulent gene exchange with other Salmonella strains harboring the gene [26,27].…”

Section: Resultsmentioning

confidence: 99%

Molecular Characterization of Salmonella Species Isolated from Animal Sources in Southern-Taraba State, North-East Nigeria

Abhadionmhen,

Anyiam,

Imarenezor

et al. 2023

IJPR

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Aims: The purpose of this research was to detect the Salmonella invA gene from chicken’s droppings in Southern Taraba, North East Nigeria. The invasive “invA” gene facilitates the invasion of epithelial cells by Salmonella isolates. Study Design: This study is experimental research involving ninety-eight (98) Salmonella isolates from chicken droppings in poultry farms and domestic chicken coops. Place and Duration of Study: This research was carried out in Wukari, Donga, Ibi and Takum towns in Southern Taraba, North East Nigeria between November 2021 and May 2023. A total of 400 (Donga,100; Wukari, 100; Ibi, 100 and Takum, 100) Chicken droppings were collected from 40 (Donga, 10; Wukari, 10; Ibi, 10 and Takum, 10) small-scale poultry farms, and 100 (Donga, 25; Wukari, 25; Ibi, 25 and Takum, 25) Chicken droppings were sampled from 20 (Donga, 5; Wukari, 5; Ibi, 5 and Takum 5) domestic chicken coops and analyzed. Methodology: Using standard microbiological techniques, Salmonella Shigella Agar (SSA) was used to confirm and validate 98 (Wukari; 36, Donga; 19, Ibi; 27, and Takum; 16) Salmonella isolates from stock culture. Isolates were further subjected to biochemical analysis. Boiling technique was used to extract the DNA of the 98 isolates. Salmonella invA gene amplification process was done using Polymerase Chain Reaction (PCR). Electrophoresis was done using agarose gel, and DNA band patterns were visualized with the aid of ultraviolent light (UV) showing different patterns and interpreted with the 100 bp DNA ladder. Results: The results of this study indicate that the invA gene was detected in 80.6% (Wukari; 86.1%, Donga; 68.5%, Ibi;85.2%, and Takum; 75%) of the 98 Salmonella isolates with an amplicon size of 284bp. This finding implies that 19 isolates which did not show the targeted band are not invasive, and even if they are, other invasive genes may be liable. Conclusion: The detection of the invasive invA invasive gene from Salmonella isolates from the fecal droppings of apparently healthy chickens from poultry farms and domestic chicken coops is really of great concern to public health. Unless a proper hygienic protocol and biosecurity measures are strictly adhered to where applicable in food processing chain, Salmonella isolates harboring the invA traits could be opportunistically invasive among immunocompromised individuals and children.

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Detection of viable Salmonella Typhi by reverse transcription-multiplex polymerase chain reaction

Cited by 8 publications

References 18 publications

Highly-sensitive detection of Salmonella typhi in clinical blood samples by magnetic nanoparticle-based enrichment and in-situ measurement of isothermal amplification of nucleic acids

Highly-sensitive detection of Salmonella typhi in clinical blood samples by magnetic nanoparticle-based enrichment and in-situ measurement of isothermal amplification of nucleic acids

Detection of Salmonella typhi Using Multiplex and Monoplex PCR in Tifoid Fever Patients

Molecular Characterization of Salmonella Species Isolated from Animal Sources in Southern-Taraba State, North-East Nigeria

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