Intergenic Spacer (Igs) Polymorphism: A New Genetic Marker for Differentiation of Toxoplasma Gondii Strains and Neospora Caninum

Fazaeli, Asghar; Carter, P. E.; Pennington, T. H.

doi:10.1645/0022-3395(2000)086[0716:isipan]2.0.co;2

Cited by 28 publications

(16 citation statements)

References 29 publications

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“…The sequence polymorphism has also been studied in a number of DNA loci of different T. gondii strains, including the genes for the heat-shock protein 70 (Lyons and Johnson, 1998), a dense granule antigen GRA6 (Fazaeli et al, 2000a) and the intergenic spacer (Fazaeli et al, 2000b). The sequence variants of T. gondii for those genes were used as markers for distinguishing T. gondii isolates from different animals and geographical localities (Høgdall et al, 2000).…”

Section: Gondiimentioning

confidence: 99%

Laboratory passage and characterization of an isolate of Toxoplasma gondii from an ocular patient in Korea

et al. 2003

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Toxoplasma gondii tachyzoites were isolated from the blood of an ocular patient, and have been successfully passaged in the laboratory, for over a year, by peritoneal inoculation in mice. The isolated parasite was designated the Korean Isolate-1 (KI-1) and its characteristics were compared with those of the RH strain, a wellknown virulent strain originating from a child who suffered from encephalitis. The morphology, pathogenicity, infectivity and cell culture characteristics of the KI-1 were similar to those of the RH strain. Both RH and KI-1 antigens were detected by an anti-T. gondii monoclonal antibody (mAb), Tg563, against the major surface protein SAG1 (30 kDa), whereas no reaction was observed against an anti-Neospora caninum mAb, 12B4. The KI-1 was confirmed as an isolate of T. gondii. A long-term laboratory maintenance and characterization of a local T. gondii isolate is reported for the first time in the Republic of Korea.

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Section: Gondiimentioning

confidence: 99%

Laboratory passage and characterization of an isolate of Toxoplasma gondii from an ocular patient in Korea

et al. 2003

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“…However, it cannot detect recombinant strains or those with unusual genotypes. 13 This problem can be alleviated by multilocus analysis of T. gondii strains with multiple markers. Furthermore, multilocus nested polymerase chain reaction (nPCR) analysis can detect as few as five parasite genomes and thus, is applicable to low-volume samples containing few parasites, which is typical of clinical specimens.…”

Section: Introductionmentioning

confidence: 99%

Direct Genotypic Characterization of Toxoplasma gondii Strains Associated with Congenital Toxoplasmosis in Tunisia (North Africa)

Boughattas¹,

Ben-Abdallah²,

Siala³

et al. 2010

The American Society of Tropical Medicine and Hygiene

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Here, we determined the Toxoplasma gondii genotype in amniotic fluid, placenta, and cerebrospinal fluid samples from 14 congenital toxoplasmosis cases in Tunisia, North Africa. Direct genotypic characterization of T. gondii strains was performed by polymerase chain reaction (PCR) amplification of six genetic markers (3'SAG2, 5' SAG2, SAG3, BTUB, GRA6, and APICO) and thereafter, was analyzed by restriction fragment-length polymorphism (RFLP). Samples were sequenced to resolve strain type whenever there were unclear enzyme digestion results. Multilocus analysis revealed that only one specimen harbored the type I allele in all studied loci, whereas the 13 others gave mixed genotype results with different alleles at different markers. Seven specimens produced RFLP profile of the recombinant strains I/III, and three produced a profile of I/II recombinant strains. The last three specimens produced complex digestion patterns. In these cases, sequence analysis revealed double peaks at known polymorphic sites, indicating the presence of multiple alleles.

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“…The ITS1 and ITS2 spacers have been used for strain differentiation and species identification in different organisms (Som et al, 2000). They have been found to be reliably valuable in more discrete phylogenetic separation of closely related species, recognition of new species, determination of conspecificity between isolates, discrimination within a species, and differentiation between piroplasm species and subspecies (Fazaeli et al, 2000; Zahler et al, 1998;Collins and Allsopp, 1999) compared to SSU rRNA.Herein, we report the development and application of a method on ITS1 and ITS2 with enhanced sensitivity and specificity in typing of T. orientalis types from field samples from suspected, infected or carrier cattle by fragment capillary electrophoresis. We further show the application of this method in quantifying (monitoring) parasites load and parasitemia in cattle experimentally infected with T. orientalis types.…”

mentioning

confidence: 99%

Rapid discrimination and quantification of Theileria orientalis types using ribosomal DNA internal transcribed spacers

Kamau

Salim

Yokoyama

et al. 2011

Infection, Genetics and Evolution

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We report the population structure analysis of Theileria orientalis types (Ikeda, Buffeli and Chitose), the causative agent of theileriosis in cattle and its cohorts, using ITS1 and ITS2 spacers by fragment genotyping. We utilized primers flanking the two ribosomal RNA internal transcribed spacers (ITS1 and ITS2). Due to varying degrees of sequence polymorphism in the ITS regions found within and between species, we exploited the insertions and or deletions in these regions which resulted in different fragment sizes. On the basis of fragment size polymorphism, we were able to discriminate the three commonly found types of Theileria orientalis. ITS1 was capable of discriminating all three types (Ikeda-251bp, Chitose-274bp and Buffeli-269bp) in one single reaction by fragment genotyping. However, using ITS2, Ikeda (133-bp) a more pathogenic type was distinguishable from Buffeli/Chitose (139-bp). In addition, we quantified parasite load in experimental animals using ITS1. When compared with previous PCR detection method, ITS1 and ITS2 genotyping were found to be more sensitive methods with high specificity in population analysis and can be deployed in molecular epidemiology studies.Keywords: Ribosomal RNA intergenic spacer; Theileria orientalis types; genotyping, MPSP INTRODUCTIONAsia-Pacific theileriosis is caused by a parasite known variously as T. sergenti or T.orientalis; although the name T. sergenti is now considered redundant (C. Sugimoto, pers.Comm.). Theileriosis due to T. orientalis is prevalent throughout Japan, Korea and parts of Russia and China (Chae et al., 1999;Fujisaki et al., 1991; Kawazu et al., 1992a, b;Onuma et al., 1998) and has been reported to occur in Australia (Callow, 1984;Kamau et al., 2010). The distribution of T. orientalis-buffeli appears to be worldwide, but is less frequently associated with clinical disease although some fatal cases have been reported to occur in America (Chae et al., 1999;Stockham et al., 2000; Cossio-Buyagur et al., 2002). Its transmitted by the three-hosts tick vector Haemaphysalis longicornis, and characterized clinically by anemia and weakness, leading to death, especially in animals experiencing metabolic stress such as parturition. There are eight genetic types (genotypes) currently comprising the T. orientalis complex. These types include type1 (Chitose), type2 (Ikeda), type3 (Buffeli) and type-4-8 yet to be classified with taxonomically (Kim et al., 1998;Ota et al., 2009). Mixed infection with different types of T. orientalis in cattle are common in Japan (Kim et al., 2004;Kubota et al., 1995), Korea (Baek et al., 2002) and Thailand (Sarataphan et al., 2003) but their clinical significance remain unclear. According to Shimizu et al., (1992;, at least one to two billion yen (one to two million US$) per year is spent to fight theileria infections in Japan.Diagnosis of a variety of disease syndromes caused by the T. orientalis parasites is principally based on clinical signs, knowledge of the disease, tick-vector distribution, and identification of...

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Intergenic Spacer (Igs) Polymorphism: A New Genetic Marker for Differentiation of Toxoplasma Gondii Strains and Neospora Caninum

Cited by 28 publications

References 29 publications

Laboratory passage and characterization of an isolate of Toxoplasma gondii from an ocular patient in Korea

Laboratory passage and characterization of an isolate of Toxoplasma gondii from an ocular patient in Korea

Direct Genotypic Characterization of Toxoplasma gondii Strains Associated with Congenital Toxoplasmosis in Tunisia (North Africa)

Rapid discrimination and quantification of Theileria orientalis types using ribosomal DNA internal transcribed spacers

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