共查询到20条相似文献,搜索用时 15 毫秒
1.
The sensitivity and accuracy of molecular diagnosis of Salmonella from meat and poultry products using polymerase chain reaction (PCR) was compared with conventional microbiological methods. A total of 212 samples representing the most frequently used fresh and frozen meat and poultry products (whole, cut, ground, and processed) were collected from different locations within the city of Irbid. DNA was extracted directly from each food sample and amplified using Salmonella-specific primers. Samples were also analysed using conventional microbiological methods for the presence of Salmonella spp. Results showed that Salmonella was detected in 185 samples out of 212 (87%) by PCR technique, while 172 (81%) samples were detected Salmonella positive by conventional microbiological methods. On the other hand, 27 (12.7%) samples were negative by PCR and 40 (18.8%) samples were negative by conventional microbiological methods. PCR assay proved to be an effective method for Salmonella detection in meat and poultry products with high specificity and sensitivity and more importantly a less time-consuming procedure. Using PCR, Salmonella spp. detection could be achieved within 24–36 h compared to 3–8 days for the conventional microbiological methods. 相似文献
2.
Edel O'Regan Evonne McCabe Catherine Burgess Sheila McGuinness Thomas Barry Geraldine Duffy Paul Whyte Séamus Fanning 《BMC microbiology》2008,8(1):156
Background
A real-time multiplex PCR assay was developed for the detection of multiple Salmonella serotypes in chicken samples. Poultry-associated serotypes detected in the assay include Enteritidis, Gallinarum, Typhimurium, Kentucky and Dublin. The traditional cultural method according to EN ISO 6579:2002 for the detection of Salmonella in food was performed in parallel. The real-time PCR based method comprised a pre-enrichment step in Buffered Peptone Water (BPW) overnight, followed by a shortened selective enrichment in Rappaport Vasilliadis Soya Broth (RVS) for 6 hours and subsequent DNA extraction. 相似文献3.
Yang Hong Tongrui Liu Margie D Lee Charles L Hofacre Marie Maier David G White Sherry Ayers Lihua Wang Roy Berghaus John J Maurer 《BMC microbiology》2008,8(1):178
Background
Classical Salmonella serotyping is an expensive and time consuming process that requires implementing a battery of O and H antisera to detect 2,541 different Salmonella enterica serovars. For these reasons, we developed a rapid multiplex polymerase chain reaction (PCR)-based typing scheme to screen for the prevalent S. enterica serovars Enteritidis, Hadar, Heidelberg, and Typhimurium. 相似文献4.
Background
Salmonella enterica is a facultative intracellular pathogen that replicates within a membrane-bound compartment termed Salmonella containing vacuole (SCV). The biogenesis of SCV requires Salmonella type III protein secretion/translocation system and their effector proteins which are translocated into host cells to exploit the vesicle trafficking pathways. SseF is one of these effectors required for SCV formation and Intracellular Salmonella replication through unknown mechanisms. 相似文献5.
R.P. Smith V. Andres F. Martelli B. Gosling F. Marco‐Jimenez K. Vaughan M. Tchorzewska R. Davies 《Journal of applied microbiology》2018,124(1):274-285
Aims
The control of Salmonella in pig production is necessary for public and animal health, and vaccination was evaluated as a strategy to decrease pig prevalence.Methods and Results
The study examined the efficacy of a live Salmonella Typhimurium vaccine, administered to sows on eight commercial farrow‐to‐finish herds experiencing clinical salmonellosis or Salmonella carriage associated with S. Typhimurium or its monophasic variants. Results of longitudinal Salmonella sampling were compared against eight similarly selected and studied control farms. At the last visit (~14 months after the start of vaccination), when all finishing stock had been born to vaccinated sows, both faecal shedding and environmental prevalence of Salmonella substantially declined on the majority of vaccinated farms in comparison to the controls. A higher proportion of vaccine farms resolved clinical salmonellosis than controls. However, Salmonella counts in positive faeces samples were similar between nonvaccinated and vaccinated herds.Conclusions
The results suggest that maternal vaccination is a suitable option for a Salmonella Typhimurium reduction strategy in farrow‐to‐finish pig herds.Significance and Impact of the Study
Salmonella vaccines have the potential to reduce the prevalence of Salmonella in pigs and result in a reduction of human cases attributed to pork. 相似文献6.
Chai-Hoon Khoo Yoke-Kqueen Cheah Learn-Han Lee Jiun-Horng Sim Noorzaleha Awang Salleh Shiran Mohd Sidik Son Radu Sabrina Sukardi 《Antonie van Leeuwenhoek》2009,96(4):441-457
The increased occurrence of Salmonella occurrence in local indigenous vegetables and poultry meat can be a potential health hazards. This study is aimed to detect
the prevalence of twenty different virulence factors among Salmonella enterica strains isolated from poultry and local indigenous vegetables in Malaysia via an optimized, rapid and specific multiplex
PCR assay. The assay encompasses a total of 19 Salmonella pathogenicity islands genes and a quorum sensing gene (sdiA) in three multiplex reaction sets. A total of 114 Salmonella
enterica isolates belonging to 38 different serovars were tested. Each isolate in under this study was found to possess up to 70%
of the virulence genes tested and exhibited variable pathogenicity gene patterns. Reproducibility of the multiplex PCR assay
was found to be 100% and the detection limit of the optimized multiplex PCR was tested with lowest detectable concentration
of DNA 0.8 pg μl−1. This study demonstrated various Salmonella pathogenicity island virulence gene patterns even within the same serovar. This sets of multiplex PCR system provide a fast
and reliable typing approach based on Salmonella pathogenicity islands, thus enabling an effective monitoring of emerging pathogenic Salmonella strains as an additional tool in Salmonella surveillance studies. 相似文献
7.
Tina Mygind Svend Birkelund Niels H Birkebæk Lars Østergaard Jørgen Skov Jensen Gunna Christiansen 《BMC microbiology》2002,2(1):17-8
Background
Chlamydia pneumoniae infection has been detected by serological methods, but PCR is gaining more interest. A number of different PCR assays have been developed and some are used in combination with serology for diagnosis. Real-time PCR could be an attractive new PCR method; therefore it must be evaluated and compared to conventional PCR methods. 相似文献8.
9.
Ivan Rychlik Daniela Karasova Alena Sebkova Jiri Volf Frantisek Sisak Hana Havlickova Vladimir Kummer Ariel Imre Annamaria Szmolka Bela Nagy 《BMC microbiology》2009,9(1):268
Background
Salmonella is a highly successful parasite of reptiles, birds and mammals. Its ability to infect and colonise such a broad range of hosts coincided with the introduction of new genetic determinants, among them 5 major pathogeniCity islands (SPI1-5), into the Salmonella genome. However, only limited information is available on how each of these pathogeniCity islands influences the ability of Salmonella to infect chickens. In this study, we therefore constructed Salmonella Enteritidis mutants with each SPI deleted separately, with single individual SPIs (i.e. with the remaining four deleted) and a mutant with all 5 SPIs deleted, and assessed their virulence in one-day-old chickens, together with the innate immune response of this host. 相似文献10.
The purpose of the present study was to investigate the application of various sample preparation methods (cell washing before lysis, purification of DNA using phenol extraction method, immunomagnetic separation-IMS) for the final PCR identification of Salmonellacells. The presence of PCR inhibitors in processed food products (milk powder and dried eggs) can be the cause of false-negative results in PCR without IMS of target cells. It was also demonstrated that IMS-PCR was successfully used for identification and quick confirmation of untypical Salmonella strains isolated from human stool samples and rabbit meat. However, IMS cannot eliminate intracellular PCR inhibitors present in immunoseparated Salmonella cells. These inhibitors must be taken into consideration in evaluation of PCR procedure. 相似文献
11.
Background
SpiC encoded within Salmonella pathogeniCity island 2 on the Salmonella enterica serovar Typhimurium chromosome is required for survival within macrophages and systemic infection in mice. Additionally, SpiC contributes to Salmonella-induced activation of the signal transduction pathways in macrophages by affecting the expression of FliC, a component of flagella filaments. Here, we show the contribution of SpiC in flagellum synthesis. 相似文献12.
McCabe EM Burgess CM Walsh D O'Regan E McGuinness S Barry T Fanning S Duffy G 《Journal of microbiological methods》2011,84(1):19-26
In Europe, alternative methods for the detection of food-borne pathogens can be used instead of the standard ISO/CEN reference protocol, if validated according to the protocol outlined in ISO 16140, 2003. In this study, the performance of two novel methods for the detection of Salmonella sp. using real-time PCR technology in tandem with an adapted two-step enrichment protocol were assessed and validated against a reference culture method, ISO 6579, 2004. The DNA and RNA real-time PCR assays amplified a 270 bp region of the hilA gene of Salmonella enterica serovars, and incorporated an internal amplification control (IAC) which was co-amplified with the hilA gene to monitor potential PCR inhibitors and ensure successful amplification. The inclusivity and exclusivity of the hilA primer set was examined for both the DNA and RNA methods and detected the 30 S. enterica serovars but not the 30 non-salmonellae strains. The inoculation of meat carcass swabs with five different S. enterica serovars at five different inocula, indicated both PCR methods were able to detect between 1 and 10 CFU per carcass swab. The real-time DNA PCR assay performed as well as the traditional cultural method in detecting Salmonella sp. in artificially contaminated salad, chocolate, fish and cheese samples. The relative accuracy, relative sensitivity and relative specificity of the DNA PCR real-time method were determined to be 98.5, 98.1 and 100%, respectively. The DNA method was further validated in a collaborative inter-laboratory trial according to ISO 16140, 2003. The validated methods provide an accurate means for the rapid detection and tracking of S. enterica serovars giving equivalent results to the standard method within three days, thus providing an alternative testing method to the reference microbiological method. The real-time PCR methodology not only offers significant time-saving advantages compared to traditional methods, it can also be applied to a wide range of samples types. 相似文献
13.
Pietro Calderini Marta Magi Simona Gabrielli Alberto Brozzi Susanna Kumlien Goffredo Grifoni Albertina Iori Gabriella Cancrini 《BMC veterinary research》2009,5(1):1-6
Background
Feed contaminated with Salmonella spp. constitutes a risk of Salmonella infections in animals, and subsequently in the consumers of animal products. Salmonella are occasionally isolated from the feed factory environment and some clones of Salmonella persist in the factory environment for several years. One hypothesis is that biofilm formation facilitates persistence by protecting bacteria against environmental stress, e.g. disinfection. The aim of this study was to investigate the biofilm forming potential of Salmonella strains from feed- and fishmeal factories. The study included 111 Salmonella strains isolated from Norwegian feed and fish meal factories in the period 1991–2006 of serovar Agona, serovar Montevideo, serovar Senftenberg and serovar Typhimurium. 相似文献14.
Susanna Kangas Tapani Lyytikäinen Jukka Peltola Jukka Ranta Riitta Maijala 《Acta veterinaria Scandinavica》2007,49(1):35
Background
Costs and benefits of two Salmonella control policies for broiler production were described and compared. The control options were the Zoonosis Directive 92/117/EC and the more intense strategy, the Finnish Salmonella Control Programme (FSCP). 相似文献15.
Burkhard Malorny Elisa Paccassoni Patrick Fach Cornelia Bunge Annett Martin Reiner Helmuth 《Applied microbiology》2004,70(12):7046-7052
A robust 5′ nuclease (TaqMan) real-time PCR was developed and validated in-house for the specific detection of Salmonella in food. The assay used specifically designed primers and a probe target within the ttrRSBCA locus, which is located near the Salmonella pathogenicity island 2 at centisome 30.5. It is required for tetrathionate respiration in Salmonella. The assay correctly identified all 110 Salmonella strains and 87 non-Salmonella strains tested. An internal amplification control, which is coamplified with the same primers as the Salmonella DNA, was also included in the assay. The detection probabilities were 70% when a Salmonella cell suspension containing 103 CFU/ml was used as a template in the PCR (5 CFU per reaction) and 100% when a suspension of 104 CFU/ml was used. A pre-PCR sample preparation protocol including a preenrichment step in buffered peptone water followed by DNA extraction-purification was applied when 110 various food samples (chicken rinses, minced meat, fish, and raw milk) were investigated for Salmonella. The diagnostic accuracy was shown to be 100% compared to the traditional culture method. The overall analysis time of the PCR method was approximately 24 h, in contrast to 4 to 5 days of analysis time for the traditional culture method. This methodology can contribute to meeting the increasing demand of quality assurance laboratories for standard diagnostic methods. Studies are planned to assess the interlaboratory performance of this diagnostic PCR method. 相似文献
16.
Arvind K Awasthi GM Nagaraja GV Naik Sriramana Kanginakudru K Thangavelu Javaregowda Nagaraju 《BMC genetics》2004,5(1):1
Background
The genus Morus, known as mulberry, is a dioecious and cross-pollinating plant that is the sole food for the domesticated silkworm, Bombyx mori. Traditional methods using morphological traits for classification are largely unsuccessful in establishing the diversity and relationships among different mulberry species because of environmental influence on traits of interest. As a more robust alternative, PCR based marker assays including RAPD and ISSR were employed to study the genetic diversity and interrelationships among twelve domesticated and three wild mulberry species. 相似文献17.
Tristano Bacchetti De Gregoris Marco Borra Elio Biffali Thomas Bekel J Grant Burgess Richard R Kirby Anthony S Clare 《BMC molecular biology》2009,10(1):62
Background
Balanus amphitrite is a barnacle commonly used in biofouling research. Although many aspects of its biology have been elucidated, the lack of genetic information is impeding a molecular understanding of its life cycle. As part of a wider multidisciplinary approach to reveal the biogenic cues influencing barnacle settlement and metamorphosis, we have sequenced and annotated the first cDNA library for B. amphitrite. We also present a systematic validation of potential reference genes for normalization of quantitative real-time PCR (qRT-PCR) data obtained from different developmental stages of this animal. 相似文献18.
Päivikki Perko-Mäkelä Pauliina Isohanni Marianne Katzav Marianne Lund Marja-Liisa Hänninen Ulrike Lyhs 《Acta veterinaria Scandinavica》2009,51(1):18
Background
Campylobacter is the most common cause of bacterial enteritis worldwide. Handling and eating of contaminated poultry meat has considered as one of the risk factors for human campylobacteriosis. Campylobacter contamination can occur at all stages of a poultry production cycle. The objective of this study was to determine the occurrence of Campylobacter during a complete turkey production cycle which lasts for 1,5 years of time. For detection of Campylobacter, a conventional culture method was compared with a PCR method. Campylobacter isolates from different types of samples have been identified to the species level by a multiplex PCR assay. 相似文献19.
20.
Alberto Biscontin Silvia Casara Stefano Cagnin Lucia Tombolan Angelo Rosolen Gerolamo Lanfranchi Cristiano De Pittà 《BMC molecular biology》2010,11(1):44