Optimization and evaluation of a multiplex PCR for simultaneous detection of prominent foodborne pathogens of Enterobacteriaceae

Babu, Litty; Reddy, Prakash Narayana; Murali, H. S.; Batra, Harsh Vardhan

doi:10.1007/s13213-013-0622-0

Cited by 16 publications

(12 citation statements)

References 23 publications

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“…Detection of very low levels of bacterial contamination in food necessitates that these samples to be cultured for a few hours in peptone water for providing conditions for growth and multiplication of bacterial pathogens to a detectable level, dilution of inhibitory substances present in food and dilution of dead target cells, which provides some assurance that the detected DNA belongs from viable target cells. The multiplex PCR reported in our study had a reasonably high level of sensitivity in experimentally spiked chicken pulav samples and able to detect as low as 10 1 and 10 2 organisms per ml of Shigella and Salmonella enterica respectively following 5 h enrichment in peptone water thus the sensitivity was much better when compared to other reports (Babu et al , 2013; Ojha et al , 2013). According to Ohja et al (2013) detection limit of mpcr after preincubation in GNB (Gram Negative Broth) was 5 × 10 4 cells and according to Babu et al (2013) mpcr detection limit was 10 cfu /PCR after 10 h of incubation in BHI broth.…”

Section: Discussionsupporting

confidence: 43%

“…The multiplex PCR reported in our study had a reasonably high level of sensitivity in experimentally spiked chicken pulav samples and able to detect as low as 10 1 and 10 2 organisms per ml of Shigella and Salmonella enterica respectively following 5 h enrichment in peptone water thus the sensitivity was much better when compared to other reports (Babu et al , 2013; Ojha et al , 2013). According to Ohja et al (2013) detection limit of mpcr after preincubation in GNB (Gram Negative Broth) was 5 × 10 4 cells and according to Babu et al (2013) mpcr detection limit was 10 cfu /PCR after 10 h of incubation in BHI broth.…”

Section: Discussionsupporting

confidence: 43%

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A novel multiplex PCR for the simultaneous detection of Salmonella enterica and Shigella species

Radhika¹,

Majumder²,

Murali³

et al. 2014

Braz. J. Microbiol.

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Salmonella enterica and Shigella species are commonly associated with food and water borne infections leading to gastrointestinal diseases. The present work was undertaken to develop a sensitive and reliable PCR based detection system for simultaneous detection of Salmonella enterica and Shigella at species level. For this the conserved regions of specific genes namely ipaH1, ipaH, wbgZ, wzy and invA were targeted for detection of Shigella genus, S. flexneri, S. sonnei, S. boydii and Salmonella enterica respectively along with an internal amplification control (IAC). The results showed that twenty Salmonella and eleven Shigella spp., were accurately identified by the assay without showing non-specificity against closely related other Enterobacteriaceae organisms and also against other pathogens. Further evaluation of multiplex PCR was undertaken on 50 natural samples of chicken, eggs and poultry litter and results compared with conventional culture isolation and identification procedure. The multiplex PCR identified the presence of Salmonella and Shigella strains with a short pre-enrichment step of 5 h in peptone water and the same samples were processed by conventional procedures for comparison. Therefore, this reported multiplex PCR can serve as an alternative to the tedious time-consuming procedure of culture and identification in food safety laboratories.

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Section: Discussionsupporting

confidence: 43%

Section: Discussionsupporting

confidence: 43%

A novel multiplex PCR for the simultaneous detection of Salmonella enterica and Shigella species

Radhika¹,

Majumder²,

Murali³

et al. 2014

Braz. J. Microbiol.

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“…Whole-genomic DNA extraction was performed by boiling lysis method, preparing the cell suspension of purely isolated bacterial colonies, as performed previously [19]. Already designed conserved regions, specific blaOXA, blaSHV, and blaTEM primers were optimized for multiplex PCR [20,21].…”

Section: Bla Oxa Blashv and Blatem Multiplex Pcrmentioning

confidence: 99%

blaOXA, blaSHV-, and blaTEM- extended-spectrum β-lactamases in Gram-negative strains from burn patients in Lahore, Pakistan

Haider

Ain

Abrar

et al. 2020

J Infect Dev Ctries

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Introduction: Gram-negative bacterial pathogens are associated with complications of post-burn infections that showed significant association with different genotypic variants of extended-spectrum β-lactamases. In this study, we aimed to determine the distribution of ESBL gene variants among bacterial pathogens from post-burn infections. Methodology: Cephalosporin-resistant isolates were collected from Jinnah Burn and Reconstructive Surgery Centre, Lahore. Phenotypic testing: double-disk synergy test, combination disk test, multiplex PCR for blaOXA, blaTEM, and blaSHV genes were performed. Results: Of the 53.5% cephalosporin-resistant isolates, 50.7% were community-acquired and 49.3% were nosocomial pathogens. Seventy-two percent of post-burn infections were found in males (p-value = 0.92, OR = 1.04). The age of burn victims ranged from 4 to 85 years (mean = 28.9, SD = 15.6). Pseudomonas aeruginosa was most predominant at a rate of 49%. There were 83.3% multidrug-resistant isolates, which showed susceptibility to meropenem, imipenem, and amikacin in 28.7%, 25.3%, and 26% respectively. In phenotypic tests 16% ESBLs detected by the DDST and 14% confirmed by the CDT. Molecular detection proved effective for the detection of 79.7% blaTEM, 37.7% blaOXA, and 18.8% blaSHV isolates. blaTEM genes were confirmed in 18.1% CDT-positive isolates, with 62.6% diagnostic accuracy (95% CI = 54.7-70) and 88% specificity (95% CI = 80.4-93.4). Conclusions: The antimicrobial resistance associated with the ESBL-producing Pseudomonas spp. and Enterobacteriaceae is becoming a challenge for the treatment and survival of burn patients. The high rate blaTEM, blaOXA, and blaSHV genes confirm the need to improve the management of burn patients in order to prevent post-burn infections.

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“…Foram então coletadas alíquotas de 1,0 mL do caldo GN para a extração do DNA total. (Okuno et al,2018;Choi et al, 2013;Babu et al,2013)…”

Section: Preparo Das Amostrasunclassified

Vegetais minimamente processados comercializados no município do Rio de Janeiro: pesquisa de Stenotrophomonas maltophilia, Pseudomonas aeruginosa e de Salmonella spp.

Nilo

Silva²,

Marin³

2022

RSD

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Introdução: Os vegetais minimamente processados são cada vez mais reconhecidos como importantes veículos para a transmissão de patógenos humanos. A importância clínica de S. maltophilia, P. aeruginosa e Salmonela spp. tem sido reconhecida por serem relevantes microrganismos patógenos oportunistas. Sabe-se que a alimentação é a principal forma para a entrada de bactérias no trato gastrointestinal humano, e as bactérias com genes de resistência a antibióticos ingeridas poderiam transferir tais genes às bactérias patogênicas ou oportunistas do trato gastrointestinal. Objetivo: O objetivo do trabalho foi pesquisar a presença de Stenotrophomonas maltophilia, Pseudomonas aeruginosa e Salmonella spp. em vegetais minimamente processados e comercializados no município do Rio de Janeiro através do método de reação em cadeia da polimerase. Metodologia: Foram coletadas 30 amostras de vegetais minimamente processados de diferentes tipos, no período de março de 2017 a março de 2018. Para identificar a presença das bactérias nas amostras foi realizada a extração de DNA com posterior análise de Reação em Cadeia da Polimerase e visualização no gel de agarose. Resultados e Discussão: No presente estudo em nenhuma das 30 amostras analisadas por PCR foi encontrada a presença de Stenotrophomonas maltophilia, Pseudomonas aeruginosa e Salmonella spp. Conclusão: O resultado encontrado não descarta a presença dessas bactérias em vegetais minimamente processados, que já foram encontrados em diversos estudos. Os vegetais minimamente processados possuem uma grande microbiota com diferentes gêneros de bactérias e essas com resistência aos antibióticos, podendo se tornar um risco aos consumidores.

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Optimization and evaluation of a multiplex PCR for simultaneous detection of prominent foodborne pathogens of Enterobacteriaceae

Cited by 16 publications

References 23 publications

A novel multiplex PCR for the simultaneous detection of Salmonella enterica and Shigella species

A novel multiplex PCR for the simultaneous detection of Salmonella enterica and Shigella species

blaOXA, blaSHV-, and blaTEM- extended-spectrum β-lactamases in Gram-negative strains from burn patients in Lahore, Pakistan

Vegetais minimamente processados comercializados no município do Rio de Janeiro: pesquisa de Stenotrophomonas maltophilia, Pseudomonas aeruginosa e de Salmonella spp.

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