Genotyping and detection of multiple infections of Toxoplasma gondii using Pyrosequencing

Sreekumar, C.; Hill, Dolores E.; Miska, Katarzyna B.; Vianna, M.C.B.; Yan, Liying; Myers, R.; Dubey, J. P.

doi:10.1016/j.ijpara.2005.03.017

Cited by 30 publications

(24 citation statements)

References 19 publications

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“…Emphasis will be given in the future to establishing high throughput molecular assays such as pyrosequencing, as well as their field applicability. Pyrosequencing techniques have the added advantage of allowing the simultaneous detection of multiple species/genotypes in a single sample [10,11,12]. Multiplex PCR (mPCR) also enables the amplification of more than one target of interest in a PCR by using multiple primer pairs and producing amplicons of different size [13].…”

Section: Molecular Toolsmentioning

confidence: 99%

The molecular epidemiology of parasite infections: Tools and applications

Lymbery

Thompson

2012

Molecular and Biochemical Parasitology

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Molecular epidemiology, broadly defined, is the application of molecular genetic techniques to the dynamics of disease in a population. In this review, we briefly describe molecular and analytical tools available for molecular epidemiological studies and then provide an overview of how they can be applied to better understand parasitic disease. A range of new molecular tools have been developed in recent years, allowing for the direct examination of parasites from clinical or environmental samples, and providing access to relatively cheap, rapid, high throughput molecular assays. At the same time, new analytical approaches, in particular those derived from coalescent theory, have been developed to provide more robust estimates of evolutionary processes and demographic parameters from multilocus, genotypic data. To date, the primary application of molecular epidemiology has been to provide specific and sensitive identification of parasites and to resolve taxonomic issues, particularly at the species level and below. Population genetic studies have also been used to determine the extent of genetic diversity among populations of parasites and the degree to which this diversity is associated with different host cycles or epidemiologically important phenotypes. Many of these studies have also shed new light on transmission cycles of parasites, particularly the extent to which zoonotic transmission occurs, and on the prevalence and importance of mixed infections with different parasite species or intraspecific variants (polyparasitism). A major challenge, and one which is now being addressed by an increasing number of studies, is to find and utilise genetic markers for complex traits of epidemiological significance, such as drug resistance, zoonotic potential and virulence.

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Section: Molecular Toolsmentioning

confidence: 99%

The molecular epidemiology of parasite infections: Tools and applications

Lymbery

Thompson

2012

Molecular and Biochemical Parasitology

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“…Humans become infected by eating undercooked meat from infected animals or ingestion of food and water contaminated with oocysts. The infection causes mental retardation and loss of vision in children and abortion in pregnant women and livestock (Sreekumar et al, 2005;Shaapan, 2015). Once the T. gondii tachyzoites infective stages were invade host they multiply by endodyogony in host tissue and might form pseudo cysts filled with crescent or banana shaped centrally located nucleus and when cysts disintegrated it spread by host blood lead to parasitemia and invade other cells.…”

Section: Introductionmentioning

confidence: 99%

Detection of Toxoplasma gondii (Apicomplexa: Sarcocystidae) in the Brown Dog Tick Rhipicephalus sanguineus (Acari: Ixodidae) Fed on Infected Rabbits

Abdel-Shafy¹,

Shaapan²,

Abdelrahma³

et al. 2015

Research J. of Parasitology

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The brown dog tick Rhipicephalus sanguineus is a three-host tick that feeds primarily on dog and occasionally on other hosts, including human. Toxoplasmosis is generally considered the most important disease that causing abortion of both pregnant women and different female animals throughout the world. Therefore, the aim of this study was to determine the ability of the brown dog tick R. sanguineus to acquire Toxoplasma infection through feeding its larvae on experimentally infected rabbits with T. gondii. The R. sanguineous larvae were feed on rabbits experimentally infected with locally isolated virulent Toxoplasma gondii strain. Nymphs moulted from these larvae were investigated to detect the presence of T. gondii specific B1 gene DNA using PCR at 193 bp. Histological examination for liver, lung and heart of experimentally infected rabbits was performed to confirm the infection of animals with T. gondii. The histopathological examination of infected rabbit tissues (heart, lung liver) revealed infiltration of T. gondii tachyzoites and polymorph nuclear inflammatory cells in addition the presence of different tissue alterations and degeneration other than that in normally histological tissues of non-infected rabbits. The nymphs of R. sanguineus were free from T. gondii. Further investigations are needed on the other developmental stages of R. sanguineus to ensure the ability of this tick species in transmission of T. gondii.

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“…Humans become infected by eating undercooked meat from infected animals and food and water contaminated with oocysts (Sreekumar et al, 2005). Besides vertical infection during pregnancy, humans and animals can get infected either by oral uptake of sporulated Toxoplasma oocysts or by ingestion of tissue cysts upon consumption of raw or undercooked meat of infected slaughtered animals (Zhou et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Comparison of PCR and Serologic Survey for Diagnosis of Toxoplasmosis in Sheep

Shaapan¹,

Kandil²,

Nassar³

2015

Research J. of Parasitology

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Comparative diagnosis of Toxoplasma gondii infection in sheep were carried out with PCR and some serological assays using 200 blood and serum samples collected from sheep of different ages and sexes slaughtered in the main abattoir in Cairo and Giza, Egypt. PCR showed the higher prevalence of toxoplasmosis (48.5%) followed by the Modified Agglutination Test (MAT) (45.5%) and the Latex Agglutination Test (LAT) (41.0%), while the lowest prevalence was detected with the Indirect Hem-Agglutination Test (IHAT) (38.5%). When the data of the serological tests were compared with that of the PCR, as a reference test for toxoplasmosis, MAT had the highest sensitivity (95.9%) followed by LAT (90.7%) and the lowest sensitivity by IHAT (80.4%). On the other hand IHAT had the highest specificity (91.3%) followed by MAT (88.3%) and the lowest specificity was by LAT (84.5%). The present study adopt PCR and serological survey of T. gondii antibodies in sheep by using more sensitive and specific antigens prepared from isolated T. gondii local strain and the obtained results suggests that MAT alone or with LAT can be used as a highly sensitive screening test followed by PCR as a specific confirmatory test for diagnosis of toxoplasmosis in sheep. Consequently, the high prevalence of sheep toxoplasmosis detected by this study scopes the public health significance of sheep's meat as source of human infection.

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Genotyping and detection of multiple infections of Toxoplasma gondii using Pyrosequencing

Cited by 30 publications

References 19 publications

The molecular epidemiology of parasite infections: Tools and applications

The molecular epidemiology of parasite infections: Tools and applications

Detection of Toxoplasma gondii (Apicomplexa: Sarcocystidae) in the Brown Dog Tick Rhipicephalus sanguineus (Acari: Ixodidae) Fed on Infected Rabbits

Comparison of PCR and Serologic Survey for Diagnosis of Toxoplasmosis in Sheep

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