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

Chai, Jong-Yil; Lin, Ang; Shin, Eun‐Hee; Oh, Myoung Don; Han, Eun‐Taek; Nam, Ho-Woo; Lee, Soon-Hyung

doi:10.3347/kjp.2003.41.3.147

Cited by 28 publications

(32 citation statements)

References 23 publications

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“…In Korea, the positive rate for anti-Toxoplasma antibodies is approximately 7% [13,14] and several clinical cases have been reported [15], but KI-1 is the only strain that has been maintained [6]. The virulence and tissue culture characteristics of this unique isolate resemble those of type I strains [7], but there is no information about its proteomic characteristics.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, T. gondii tachyzoites, designated as Korean Isolate-1 (KI-1), were successfully maintained in the laboratory [6]. They proved to be highly virulent when inoculated into BALB/c mice and cultured in sarcoma 180 cells [6], and were assigned to the genotype I clonal lineage using PCR-based restriction fragment length polymorphism (PCR-RFLP) analysis [7].…”

Section: Introductionmentioning

confidence: 99%

“…They proved to be highly virulent when inoculated into BALB/c mice and cultured in sarcoma 180 cells [6], and were assigned to the genotype I clonal lineage using PCR-based restriction fragment length polymorphism (PCR-RFLP) analysis [7].…”

Section: Introductionmentioning

confidence: 99%

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Proteomic Analysis ofToxoplasma gondiiKI-1 Tachyzoites

et al. 2010

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Abstract:We studied on the proteomic characteristics of Toxoplasma gondii KI-1 tachyzoites which were originally isolated from a Korean patient, and compared with those of the well-known virulent RH strain using 2-dimensional electrophoresis (2-DE), mass spectrometry, and quantitative real-time PCR. Two-dimensional separation of the total proteins isolated from KI-1 tachyzoites revealed up to 150 spots, of which 121 were consistent with those of RH tachyzoites. Of the remaining 29 spots, 14 showed greater than 5-fold difference in density between the KI-1 and RH tachyzoites at a pH of 5.0-8.0. Among the 14 spots, 5 from the KI-1 isolate and 7 from the RH strain were identified using MALDI-TOF mass spectrometry and database searches. The spots from the KI-1 tachyzoites were dense granule proteins (GRA 2, 3, 6, and 7), hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGRPTase), and uracil phosphoribosyltransferase (UPRTase). The spots from the RH strain were surface antigen 1 (SAG 1), L-lactate dehydrogenase (LDH), actin, chorismate synthase, peroximal catalase, hexokinase, bifunctional dihydrofolate reductase-thymidylate synthase (DHTR-TS), and nucleosidetriphosphatases (NTPases). Quantitative real-time PCR supported our mass spectrometric results by showing the elevated expression of the genes encoding GRA 2, 3, and 6 and UPRTase in the KI-1 tachyzoites and those encoding GRA 7, SAG 1, NTPase, and chorismate synthase in the RH tachyzoites. These observations demonstrate that the protein compositions of KI-1 and RH tachyzoites are similar but differential protein expression is involved in virulence.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Proteomic Analysis ofToxoplasma gondiiKI-1 Tachyzoites

et al. 2010

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“…In our study, brains were not examined, and thus further studies of multiple tissues in cats are needed. In addition, free-living animals, such as stray cats and wild rodents living around the urban areas, are important for determining the epidemiological significance of T. gondii infection, since cases of human toxoplasmosis have been reported in Korea [13].…”

Section: Genotype Of Toxoplasma Gondii From Blood Of Straymentioning

confidence: 99%

Genotype of Toxoplasma gondii from Blood of Stray Cats in Gyeonggi-do, Korea

Kim

Lee

et al. 2009

Korean J Parasitol

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Toxoplasma gondii is an obligate intracellular protozoan that can infect both humans and warm-blooded animals. Among them, cats are the only animal that excretes oocysts into the environment [1]. As recently reported, 47.2% out of 106 stray cats [2] and 13.2% out of 174 stray cats [3] were T. gondii positive by nested PCR assay in Korea. Stray cats have been increasing in Korea roaming residential areas, and this increased the risk of public health hazards. T. gondii strains have been classified into 3 major genotypes (I, II, and III) according to their virulence, and among them, type I strain, such as RH, has been reported to be most virulent. This study is a follow-up of our previous study [3] for genotyping of T. gondii using 23 blood DNA samples of stray cats in Korea, which presented positive reactions by B1 gene amplification.Biodiversity of T. gondii has been reported with genetic variations among isolates from different, and even the same, phenotypes [4]. Usually, T. gondii is categorized as 3 genetic types (I, II, and III) based on polymorphisms in several markers, such as SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, PK1, and Apico genes [5,6]. We chose to develop a nested-PCR method based on the polymorphic SAG2 locus, separately amplifying the 5 ′ and 3′ ends of the locus using an Accupower PCR premix Kit (Bioneer Co., Seoul, Korea), and amplifying 241 bp and 221 bp products [7,8]. This gene is ideally suited for rapid genotyping, as it contains multiple lineage-specific polymorphisms [7,9,10]. SAG2 encodes 2 separate forms of the surface tachyzoite protein (p22), that type I (high virulent) and III (avirulent) strains share the same protein alleles, while type II strains (avirulent) have a second, distinct form [5,7]. Sau3A I and Hha I restriction enzymes were used for this PCR-RFLP analysis. Digestion of the 5 ′ amplification products with Sau3A I distinguished allele 3 (type III: includes avirulent strains like VEG and CTG) from alleles 1 and 2 (type I: includes strains like RH and KI-1 which are highly virulent, and type II: includes avirulent strains like PLK and Beverley) [8]. Digestion of the 3′ amplification products with Hha I distinguished allele 2 (type II strains) from alleles 1 and 3 (type I and III strains) [7].RH and KI-1 strains as type I, and ME49 strain as type II, were used for standardization of PCR-RFLP analysis. The representative strain as type III was not prepared because genotyping could be properly discriminated using strains mentioned above. RH strain was kindly provided from Catholic Institute of Parasitic Diseases, Catholic University, and KI-1 and ME49 strains were kindly provided from Department of Parasitology and Tropical Medicine, Seoul National University College of Medicine. RH and KI-1 strains of T. gondii were maintained in BALB/c mice by intraperitoneal inoculation in our laboratory.PCR programs were as follows: The 5′ and 3′ ends of the SAG2 locus were amplified by 5 min of denaturation at 94℃. After Genotype of Toxoplasma gondii from Blood of Stray Cats in Gyeon...

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“…A drawback of this method is the need of specialized personal because of the risk of contamination by the handling of Toxoplasma, as well as the lapse of time to get the results [1][2][3][4][5].…”

Section: Isolation Of the Parasite Or Bioassaymentioning

confidence: 99%

The Laboratory Diagnosis in Toxoplasma Infection

Ramírez¹,

Orozco²,

Ramírez³

2017

Toxoplasmosis

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The diagnosis of toxoplasmosis is of great importance due to the damage caused by this parasite in immunosuppressed people or in pregnant women, the diagnosis of an active toxoplasmosis represents a sign to initiate a pharmacological treatment immediately. The diagnosis of Toxoplasma can be performed with direct methods through intraperitoneal inoculation of serum or cerebrospinal luid, in susceptible mice evaluating the survival and detection of tachyzoites of biological samples. Indirect methods detecting the IgM and IgG isotypes against Toxoplasma have been the tools mostly used and had leaded to discriminate between an active and acute, from a chronic toxoplasmosis. Molecular methods actually Toxoplasma-DNA identiication by molecular biology tests like the polymerase chain reaction (PCR) allow the direct detection of the parasite. Polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs) have been used to identify three strain linages (type I, II and III). Recently, a high-resolution melting method was described to determine the genotype of the infection by Toxoplasma gondii directly from biological samples.

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

Cited by 28 publications

References 23 publications

Proteomic Analysis ofToxoplasma gondiiKI-1 Tachyzoites

Proteomic Analysis ofToxoplasma gondiiKI-1 Tachyzoites

Genotype of Toxoplasma gondii from Blood of Stray Cats in Gyeonggi-do, Korea

The Laboratory Diagnosis in Toxoplasma Infection

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