A simple, rapid, cost-effective and sensitive method for detection of <i>Salmonella</i>
 in environmental and pecan samples

Dobhal, Shefali; Zhang, G.; Rohla, Charles T.; Smith, Michael W.; Ma, Li

doi:10.1111/jam.12583

Cited by 17 publications

(14 citation statements)

References 38 publications

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“…Several PCRbased assays have been developed for rapid detection of Salmonella sp. [8,9] . Various serotype-specific PCR have also been developed for some common serovars to reduce time and cost in processing isolates by conventional serotyping which is very much labor intensive and time-consuming [10].…”

Section: Introductionmentioning

confidence: 99%

Isolation, serotype diversity and antibiogram of Salmonella enterica isolated from different species of poultry in India

Mir

Kashyap

Maherchandani

2015

Asian Pacific Journal of Tropical Biomedicine

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A B S T R A C TObjective: To study the occurrence and serotype diversity of Salmonella isolates in different species of poultry (chicken, emu and duck) and determine their resistance pattern against various antibiotics of different classes. Methods: About 507 samples comprising 202 caecal contents and 305 fecal samples from chicken, emu and duck were processed for isolation of Salmonella enterica. Salmonellae were isolated and detected by standard protocol of ISO 6579 Amendment 1: Annex D. Genetic confirmation was also made by using 16S rRNA genus specific PCR. Serotype specific PCR was also done to detect the most common serovars viz. Salmonella Enteritidis, Salmonella Typhimurium and Salmonella Gallinarum. All obtained isolates were subjected to a set of 25 antibiotics to study their antibiogram by using Baeur-Kirby disk diffusion method. Results: Out of 507 samples processed, 32 isolates of Salmonella enterica (18 from caecal contents and 14 from faecal samples) were obtained, of which 24 belonged to 6 different serovars, 6 were untypeable and 2 were rough strains. Salmonella Enteritidis was the most predominant serotype (9), followed by Salmonella Typhimurium (5), Salmonella Virchow (4), Salmonella Gallinarum (3), Salmonella Reading (2) and Salmonella Altona (1). Antibiotic resistance pattern was maximum (100%) to oxacillin, penicillin and clindamycin, followed by ampicillin (68.75%), tetracycline (65.62%), nalidixic acid (56.25%) and colistin (46.87%). High sensitivity of isolates was recorded for chloramphenicol (96.87%) followed by meropenem (84.37%). Conclusions: Occurrence of high proportion of serovars in our study which can cause serious gastroenteritis in humans is a matter of concern. Salmonella Altona has been detected for the first time in India from poultry. This serotype is known to cause serious outbreaks of gastroenteritis in humans. Multidrug resistant isolates were recovered at high percentage which can be attributed to non-judicious use of antibiotics both in prophylaxis and treatment regimen. This observation draws serious attention as poultry serves as an important source of transmission of these multidrug resistant Salmonella serovars to humans.

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Section: Introductionmentioning

confidence: 99%

Isolation, serotype diversity and antibiogram of Salmonella enterica isolated from different species of poultry in India

Mir

Kashyap

Maherchandani

2015

Asian Pacific Journal of Tropical Biomedicine

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“…Preliminary endpoint RT-PCR was performed with positive RRV infected tissue (positive control) with and without the addition of amplification facilitators (BSA: Bovine serum albumin, and PVP: Polyvinylpyrolidone;Dobhal et al, 2014b). Endpoint RT-PCR was performed in 20 μl…”

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confidence: 99%

A simplified strategy for sensitive detection of Rose rosette virus compatible with three RT-PCR chemistries

Dobhal

Olson

Arif

et al. 2016

Journal of Virological Methods

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Rose rosette disease is a disorder associated with infection by Rose rosette virus (RRV), a pathogen of roses that causes devastating effects on most garden cultivated varieties, and the wild invasive rose especially Rosa multiflora. Reliable and sensitive detection of this disease in early phases is needed to implement proper control measures. This study assesses a single primer-set based detection method for RRV and demonstrates its application in three different chemistries: Endpoint RT-PCR, TaqMan-quantitative RT-PCR (RT-qPCR) and SYBR Green RT-qPCR with High Resolution Melting analyses. A primer set (RRV2F/2R) was designed from consensus sequences of the nucleocapsid protein gene p3 located in the RNA 3 region of RRV. The specificity of primer set RRV2F/2R was validated in silico against published GenBank sequences and in-vitro against infected plant samples and an exclusivity panel of near-neighbor and other viruses that commonly infect Rosa spp. The developed assay is sensitive with a detection limit of 1fg from infected plant tissue. Thirty rose samples from 8 different states of the United States were tested using the developed methods. The developed methods are sensitive and reliable, and can be used by diagnostic laboratories for routine testing and disease management decisions.

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“…The genus Salmonella spp. has often been reported in studies using different microbiological and molecular techniques for the identification and differentiation in samples from company and food-producing animals (Dickel et al, 2005;Malorny et al, 2007;O'Regan et al, 2008;Mcguinness et al, 2009;Omiccioli et al, 2009;Andrade et al, 2010;Löfström et al, 2010;Park et al, 2011;Barletta et al, 2013;Dobhal et al, 2014;Li et al, 2014;Ma et al, 2014). The fast diagnosis and identification of these bacteria is crucial for the control of bacterial dissemination, reduction of losses in animal production and risks to human health caused by foodborne infections (Chen et al, 2010;Cheraghchi et al, 2014).…”

Section: Resultsmentioning

confidence: 99%

“…Poultry production is often affected by acute and subclinical infections caused by enterobacteriaceae of genus Salmonella spp. and these bacteria are responsible for health impacts and large economic losses worldwide (Dobhal et al, 2014). The outcome of the disease depends on different factors, such as age of birds, genetic resistance and immunosuppressive conditions.…”

Section: Resultsmentioning

confidence: 99%

Development of a multiplex qPCR in real time for quantification and differential diagnosis ofSalmonellaGallinarum andSalmonellaPullorum

et al. 2017

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Currently there are 2659 Salmonella serovars. The host-specific biovars Salmonella Pullorum and Salmonella Gallinarum cause systemic infections in food-producing and wild birds. Fast diagnosis is crucial to control the dissemination in avian environments. The present work describes the development of a multiplex qPCR in real time using a low-cost DNA dye (SYBr Green) to identify and quantify these biovars. Primers were chosen based on genomic regions of difference (RoD) and optimized to control dimers. Primers pSGP detect both host-specific biovars but not other serovars and pSG and pSP differentiate biovars. Three amplicons showed different melting temperatures (Tm), allowing differentiation. The pSGP amplicon (97 bp) showed Tm of 78°C for both biovars. The pSG amplicon (273 bp) showed a Tm of 86.2°C for S. Gallinarum and pSP amplicon (260 bp) dissociated at 84.8°C for S. Pullorum identification. The multiplex qPCR in real time showed high sensitivity and was capable of quantifying 10-10 CFU of these biovars.

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A simple, rapid, cost-effective and sensitive method for detection of Salmonella in environmental and pecan samples

Abstract: This study provides a method with broad applicability for PCR detection of Salmonella in complex sample matrices. This method has a potential for its application in different research arenas and diagnostic laboratories.

Cited by 17 publications

References 38 publications

Isolation, serotype diversity and antibiogram of Salmonella enterica isolated from different species of poultry in India

Isolation, serotype diversity and antibiogram of Salmonella enterica isolated from different species of poultry in India

A simplified strategy for sensitive detection of Rose rosette virus compatible with three RT-PCR chemistries

Development of a multiplex qPCR in real time for quantification and differential diagnosis ofSalmonellaGallinarum andSalmonellaPullorum

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