Direct detection and genotyping of Toxoplasma gondii in meat samples using magnetic capture and PCR

Opsteegh, Marieke; Langelaar, Merel; Sprong, Hein; Hartog, Laurien den; Craeye, Stéphane De; Bokken, G.C.A.M.; Ajzenberg, Daniel; Kijlstra, Aize; Giessen, Joke van der

doi:10.1016/j.ijfoodmicro.2010.02.027

Cited by 189 publications

(151 citation statements)

References 27 publications

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“…In an attempt to increase the sensitivity of detection by PCR, a method based on sequence-specific magnetic capture of T. gondii DNA followed by DNA amplification has been developed. It allowed the testing of 100-g tissue sample homogenates, with an estimated detection limit of approximately 1 cyst per 100 g (242).…”

Section: Prevalence and Importance Of Toxoplasmosis In The World Prevmentioning

confidence: 99%

Epidemiology of and Diagnostic Strategies for Toxoplasmosis

2012

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SUMMARY The apicomplexan parasite Toxoplasma gondii was discovered a little over 100 years ago, but knowledge of its biological life cycle and its medical importance has grown in the last 40 years. This obligate intracellular parasite was identified early as a pathogen responsible for congenital infection, but its clinical expression and the importance of reactivations of infections in immunocompromised patients were recognized later, in the era of organ transplantation and HIV infection. Recent knowledge of host cell-parasite interactions and of parasite virulence has brought new insights into the comprehension of the pathophysiology of infection. In this review, we focus on epidemiological and diagnostic aspects, putting them in perspective with current knowledge of parasite genotypes. In particular, we provide critical information on diagnostic methods according to the patient's background and discuss the implementation of screening tools for congenital toxoplasmosis according to health policies.

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Section: Prevalence and Importance Of Toxoplasmosis In The World Prevmentioning

confidence: 99%

Epidemiology of and Diagnostic Strategies for Toxoplasmosis

2012

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“…This can be done on farm or at the slaughterhouse. Studies on PCR methods to detect and quantify T. gondii in meat samples have shown promise with detection sensitivities comparable to those of bioassay (Opsteegh et al, 2010a). Studies using such laboratory techniques allow epidemiological studies to be conducted to determine the seroprevalence of toxoplasmosis in ovine meat and the risks such meat poses to human health (EFSA, 2007b;Opsteegh et al, 2010b).…”

Section: At the Slaughterhousementioning

confidence: 99%

Scientific Opinion on the public health hazards to be covered by inspection of meat from sheep and goats

2013

EFSA Journal

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A risk ranking process identified Toxoplasma gondii and pathogenic verocytotoxin-producing Escherichia coli (VTEC) as the most relevant biological hazards for meat inspection of sheep and goats. As these are not detected by traditional meat inspection, a meat safety assurance system using risk-based interventions was proposed. Further studies are required on T. gondii and pathogenic VTEC. If new information confirms these hazards as a high risk to public health from meat from sheep or goats, setting targets at carcass level should be considered. Other elements of the system are risk-categorisation of flocks/herds based on improved Food Chain Information (FCI), classification of abattoirs according to their capability to reduce faecal contamination, and use of improved process hygiene criteria. It is proposed to omit palpation and incision from post-mortem inspection in animals subjected to routine slaughter. For chemical hazards, dioxins and dioxin-like polychlorinated biphenyls were ranked as being of high potential concern. Monitoring programmes for chemical hazards should be more flexible and based on the risk of occurrence, taking into account FCI, which should be expanded to reflect the SUMMARYFollowing a request from the European Commission to the European Food Safety Authority (EFSA), the Panel on Biological Hazards (BIOHAZ) was asked to deliver a Scientific Opinion on the public health hazards to be covered by the inspection of meat from sheep and goats. The Panel was supported by the EFSA Panels on Contaminants in the Food Chain (CONTAM) and Animal Health and Welfare (AHAW) in the preparation of this Opinion. Briefly, the main risks for public health that should be addressed by meat inspection were identified and ranked; the strengths and weaknesses of the current meat inspection system were evaluated; recommendations were made for inspection methods fit for the purpose of meeting the overall objectives of meat inspection for hazards currently not covered by the meat inspection system; and recommendations for adaptations of inspection methods and/or frequencies of inspections that provide an equivalent level of protection were made. In addition, the implications for animal health and animal welfare of any changes proposed to current inspection methods were assessed.Sheep and goats were considered together, unless otherwise stated. Decision trees were developed and used for priority ranking of biological and chemical hazards present in meat from sheep and goats. For biological hazards the ranking was based on the magnitude of the human health impact, the severity of the disease in humans and the evidence supporting the role of meat from sheep and goats as a risk factor for disease in humans. The assessment was focused on the public health risks that may occur through the handling, preparation for consumption and/or consumption of meat from these species. The term 'priority' was considered more appropriate than 'risk' for categorizing the biological hazards associated with meat from small rumina...

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“…The methodology for the extraction and purification of DNA from T. gondii in tissue with magnetic beads was described by Opsteegh et al [56]. Tissue homogenization is carried out in a bag filter Stomacher400 containing a lysis buffer (100 mM Tris HCl pH 8.0, 5 mM EDTA pH 8.0, 0.2% SDS, 200mMNaCl and 40mg/L Proteinase k), by reaction using 2.5 mL of lysis buffer per gram of sample for 2 minutes at high speed.…”

Section: Magnetic Beadsmentioning

confidence: 99%

“…To remove free biotin from the sample streptavidin sepharose should be added to allow the biotin precipitation and to form a pellet by centrifuging and 10 mL of the supernatant are finally used for purification process [56,57].…”

Section: Magnetic Beadsmentioning

confidence: 99%

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Toxoplasma gondii in Meat for Human Consumption – A Brief Review of the Most Described Strategies for Its Detection and Quantification

Paredes¹,

Ortega‐Pacheco²,

Rosado-Aguilar³

et al. 2016

Significance, Prevention and Control of Food Related Diseases

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Toxoplasmosis is a parasitic zoonotic disease widely distributed worldwide and is caused by the intracellular parasite Toxoplasma gondii. The definitive host of T. gondii is the domestic cat and the entire cat family, in which the sexual stages of the parasite develop. T. gondii can also infect a wide range of intermediate hosts, affecting most warm-blooded animals including humans. In humans, toxoplasmosis is usually asymptomatic in healthy individuals, but can develop lymphadenopathy and nonspecific symptomatology or even be fatal in infants with congenital toxoplasmosis and in immunocompromised patients. Transmission to humans is mainly through food, especially by eating undercooked meat or meat contaminated with tissue cysts. This has led to various public health organizations worldwide monitoring programs on T. gondii in animals intended for human consumption, especially in meat samples. One of the techniques employed in the laboratory is that based on the polymerase chain reaction and some of its variants, which have proven to be valuable tools for the detection of T. gondii in tissues for human consumption and many other types of biological samples. The development of different strategies for the molecular detection of T. gondii has led to the identification and quantification methodologies varying widely among laboratories. Therefore, this chapter reviews the main methods of extraction, purification, detection and quantification of T. gondii DNA in tissue samples from different species destined for human consumption.

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Direct detection and genotyping of Toxoplasma gondii in meat samples using magnetic capture and PCR

Cited by 189 publications

References 27 publications

Epidemiology of and Diagnostic Strategies for Toxoplasmosis

Epidemiology of and Diagnostic Strategies for Toxoplasmosis

Scientific Opinion on the public health hazards to be covered by inspection of meat from sheep and goats

Toxoplasma gondii in Meat for Human Consumption – A Brief Review of the Most Described Strategies for Its Detection and Quantification

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