Evaluation of selective and non-selective enrichment PCR procedures for Salmonella detection

Oliveira, Sı́lvia Dias de; Rodenbusch, Carla Rosane; Cé, Milene C.; Rocha, Silvio L.S.; Canal, Cláudio Wageck

doi:10.1046/j.1472-765x.2003.01294.x

Cited by 106 publications

(76 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In this investigation, Salmonella Pullorum was detected from tissue (liver and lungs) samples by PCR with the amplification of invA gene at 12 hr PI. Olivera et al (2003) and Desai et al (2005) also detected 284-bp amplicon from tissue samples in experimental infected chickens.…”

Section: Detection Of S Pullorum By Pcr: Smentioning

confidence: 95%

Experimental pathogenesis of pullorum disease in chicks by local isolate of Salmonella Pullorum in Bangladesh

Haider

Chowdhury

Ahmed

et al. 2012

J Bangladesh Agric Univ

View full text Add to dashboard Cite

This study was undertaken to observe the experimental pathogenesis of locally isolated Salmonella enterica subspecies enterica serovar Pullorum in chicks. Fifty chicks were experimentally infected by the oral route with 2 x 10 7 (CFU) units of Salmonella Pullorum organisms reconstituted in 0.5 ml of sterile phosphate buffer saline (PBS), PH 7.2 and 50 chicks were given only 0.5 ml of sterile PBS as control. Observations were made on clinical signs, gross pathology, and reisolation of S. Pullorum from different organs and blood, histopathological lesions, detection of antibody levels and detection of S. Pullorum by PCR at different time intervals of experimental period. Five birds were randomly selected and sacrificed on 6 hrs before inoculation and 6 hrs, 12 hrs, 24 hrs, 2 days, 3 days, 1 st week, 2 nd , 3 rd and 4 th weeks of post infection (PI). The clinical signs of infected chicks were depression, loss of appetite, huddled together, loss of feed and water intake, reduced mean body weights, ruffled feathers, diarrhoea, laboured breathing and pasty vent. The highest gross lesion was (84%) unabsorbed and coagulated yolk and the lowest lesion was (32%) pericarditis and necrotic foci/ nodules in heart. Microscopically, the liver showed congestion, focal necrosis with multifocal infiltration of histiocytes in liver parenchyma. Salmonella organisms were reisolated from different organs and blood at 12 hrs PI. The antibody titre increased gradually and the highest titer was 7275.717 ± 5087.24 at 4 wks PI. In rapid plate agglutination test, the positive result was found from one wk of PI with the sera of infected birds. At 12hrs PI Salmonella was detected by PCR from 20% liver and 20% lung samples of infected birds and no Salmonella was isolated from control group. The orally inoculated Salmonella Pullorum organisms produced lesions in digestive tract, invaded digestive tracts and entered to blood and seeded to different organs in different time intervals and ultimately produced clinical signs, gross and microscopic tissue lesions with immunological response.

show abstract

Section: Detection Of S Pullorum By Pcr: Smentioning

confidence: 95%

Experimental pathogenesis of pullorum disease in chicks by local isolate of Salmonella Pullorum in Bangladesh

Haider

Chowdhury

Ahmed

et al. 2012

J Bangladesh Agric Univ

View full text Add to dashboard Cite

show abstract

“…The virulence determinant genes of Salmonella spp. are associated with a combination of either chromosomal or plasmid factors [13]. These genes have a role in adhesion, invasion, and enterotoxin production [13][14][15].…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of Salmonella Enteritidis and Salmonella Typhimurium from chicken meat in Egypt

Tarabees

El-Sayed

Shawish

et al. 2017

J Infect Dev Ctries

View full text Add to dashboard Cite

Introduction: Salmonella enterica serovars Enteritidis and Typhimurium represent the major serovars associated with human salmonellosis. Contamination of meat products with these serovars is considered the main source of infection. Methodology: In this study, 100 raw chicken meat samples were investigated for the presence of Salmonella spp., which were subsequently identified based on biochemical and serological tests as well as matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) profile. Furthermore, the isolated serovars were examined using multiplex polymerase chain reaction (PCR) for the presence of virulence genes suspected to have a role in infection. Results: S. Enteritidis was isolated from two samples (2%), while S. Typhimurium was isolated from three samples (3%) of chicken meat. Of the 17 examined virulence genes using multiplex PCR, the sitC, sopB, sifA, lpfC, spaN, sipB, invA, spiA, and msgA genes were detected in S. Enteritidis. However, the sitC, iroN, sopB, sifA, lpfC, spaN, sipB, invA, and tolC genes were successfully amplified in S. Typhimurium. Conclusions: The detection of S. Enteritidis and S. Typhimurium in meat, even at low incidence, has important implications. In addition, the data presented here is the first attempt to identify a wide range of virulence genes in Egyptian Salmonella isolates recovered from meat products. A strict public health and food safety regime is urgently needed in order to decrease the human health hazard risk associated with salmonellosis.

show abstract

“…Pre-enrichment of samples is thus necessary to lower the detection limit and dilute any inhibitory substances present in the samples. Most PCR assays carried out currently include a minimum of 6 to 8 h pre-cultivation step (Ellingson et al, 2004;Fratamico, 2003;Guo et al, 2000;Myint et al, 2006;Oliveira et al, 2002;Oliveira et al, 2003;Patel et al, 2006;Wang and Yeh, 2002) that can increase the sensitivity to one digit number of CFU, which is much better than the immunoassays that preenrich the samples for 24 to 30 h but still obtain minimum detection limits of 10 3 CFU (Soumet et al, 1999(a);Liu et al, 2001;Chen and Durst, 2006). Rahn et al (1992) reported the invA primer set that was able to discriminate between Salmonella and non-Salmonella species.…”

Section: Introductionmentioning

confidence: 99%

Optimisation of the PCR-invA primers for the detection of Salmonella in drinking and surface waters following a pre-cultivation step

Moganedi¹,

Goyvaerts²,

Venter³

et al. 2009

WSA

View full text Add to dashboard Cite

A polymerase chain reaction (PCR)-based method for the detection of Salmonella species in water samples was optimised and evaluated for speed, specificity and sensitivity. Optimisation of Mg 2+ and primer concentrations and cycling parameters increased the sensitivity and limit of detection of PCR to 2.6 x 10 4 cfu/mℓ. A 6h non-selective pre-enrichment step further increased the limit of detection to 26 cfu/mℓ. Out of 14 different Salmonella strains tested, only two, Salmonella arizonae and Salmonella pullorum, did not give positive amplification results with primers homologous to a conserved region of the invA gene. When environmental and drinking waters were assessed, a non-selective pre-enrichment step was included to increase the detection efficiency of PCR. The PCR method demonstrated specificity in the presence of other competing micro-organisms as confirmed by the conventional culture method. No false positives or negatives were observed when household and environmental water samples were tested by invA-PCR analysis parallel to the culture method.

show abstract

Evaluation of selective and non-selective enrichment PCR procedures for Salmonella detection

Cited by 106 publications

References 14 publications

Experimental pathogenesis of pullorum disease in chicks by local isolate of Salmonella Pullorum in Bangladesh

Experimental pathogenesis of pullorum disease in chicks by local isolate of Salmonella Pullorum in Bangladesh

Isolation and characterization of Salmonella Enteritidis and Salmonella Typhimurium from chicken meat in Egypt

Optimisation of the PCR-invA primers for the detection of Salmonella in drinking and surface waters following a pre-cultivation step

Contact Info

Product

Resources

About