Evaluation of a real-time PCR assay based on the single-copy SAG1 gene for the detection of Toxoplasma gondii

Heng, Yu; Huang, Bin; Zhuo, Xunhui; Chen, Xueqiu; Du, Aifang

doi:10.1016/j.vetpar.2013.06.013

Cited by 33 publications

(18 citation statements)

References 13 publications

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“…Sterile water was used as a negative control, and the DNA extracted from 500 tachyzoites of the T. gondii RH strain was used as a positive control. All reactions were performed in triplicates and incubated at 95°C for 1 min, followed by 40 cycles of incubation at 95°C for 5 s, 60°C for 15 s and 72°C for 10 s [27].…”

Section: Methodsmentioning

confidence: 99%

Partial Protective Effect of Intranasal Immunization with Recombinant Toxoplasma gondii Rhoptry Protein 17 against Toxoplasmosis in Mice

et al. 2014

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Toxoplasma gondii (T. gondii) is an obligate intracellular protozoan parasite that infects a variety of mammals, including humans. An effective vaccine for this parasite is therefore needed. In this study, RH strain T. gondii rhoptry protein 17 was expressed in bacteria as a fusion with glutathione S-transferase (GST) and the recombinant proteins (rTgROP17) were purified via GST-affinity chromatography. BALB/c mice were nasally immunised with rTgROP17, and induction of immune responses and protection against chronic and lethal T. gondii infections were investigated. The results revealed that mice immunised with rTgROP17 produced high levels of specific anti-rTgROP17 IgGs and a mixed IgG1/IgG2a response of IgG2a predominance. The systemic immune response was associated with increased production of Th1 (IFN-γand IL-2) and Th2 (IL-4) cytokines, and enhanced lymphoproliferation (stimulation index, SI) in the mice immunised with rTgROP17. Strong mucosal immune responses with increased secretion of TgROP17-specific secretory IgA (SIgA) in nasal, vaginal and intestinal washes were also observed in these mice. The vaccinated mice displayed apparent protection against chronic RH strain infection as evidenced by their lower liver and brain parasite burdens (59.17% and 49.08%, respectively) than those of the controls. The vaccinated mice also exhibited significant protection against lethal infection of the virulent RH strain (survival increased by 50%) compared to the controls. Our data demonstrate that rTgROP17 can trigger strong systemic and mucosal immune responses against T. gondii and that ROP17 is a promising candidate vaccine for toxoplasmosis.

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Section: Methodsmentioning

confidence: 99%

Partial Protective Effect of Intranasal Immunization with Recombinant Toxoplasma gondii Rhoptry Protein 17 against Toxoplasmosis in Mice

et al. 2014

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“…In order to verify the accuracy of evaluating the tachyzoite load above method, the distributions of tachyzoite in the liver tissues were measured using a sensitive real-time quantitative PCR (qRT-PCR) method as previously described [32]. Briefly, genomic DNA from the purified parasites and collected liver samples (100 mg) were extracted using a UniversalGen DNA Kit (CWBIO, China) according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Intranasal Immunisation with Recombinant Toxoplasma gondii Actin Partly Protects Mice against Toxoplasmosis

et al. 2013

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Toxoplasma gondii is a ubiquitous protozoan intracellular parasite, the causative agent of toxoplasmosis, and a worldwide zoonosis for which an effective vaccine is needed. Actin is a highly conserved microfilament protein that plays an important role in the invasion of host cells by T. gondii. This study investigated the immune responses elicited by BALB/c mice after nasal immunisation with a recombinant T. gondii actin (rTgACT) and the subsequent protection against chronic and lethal T. gondii infections. We evaluated the systemic response by proliferation, cytokine and antibody measurements, and we assessed the mucosal response by examining the levels of TgACT-specific secretory IgA (SIgA) in nasal, vaginal and intestinal washes. Parasite load was assessed in the liver and brain, and the survival of mice challenged with a virulent strain was determined. The results showed that the mice immunised with rTgACT developed high levels of specific anti-rTgACT IgG titres and a mixed IgG1/IgG2a response with a predominance of IgG2a. The systemic immune response was associated with increased production of Th1 (IFN-γ and IL-2), Th2 (IL-4) and Treg (IL-10) cytokines, indicating that not only Th1-type response was induced, but also Th2- and Treg-types responses were induced, and the splenocyte stimulation index (SI) was increased in the mice immunised with rTgACT. Nasal immunisation with rTgACT led to strong mucosal immune responses, as seen by the increased secretion of SIgA in nasal, vaginal and intestinal washes. The vaccinated mice displayed significant protection against lethal infection with the virulent RH strain (survival increased by 50%), while the mice chronically infected with RH exhibited lower liver and brain parasite loads (60.05% and 49.75%, respectively) than the controls. Our data demonstrate, for the first time, that actin triggers a strong systemic and mucosal response against T. gondii. Therefore, actin may be a promising vaccine candidate against toxoplasmosis.

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“…Mouse inoculation or tissue cultures of the clinical samples may confirm the infection by parasites but they are still impractical, labor-intensive and require several days to obtain results [8,9]. The acute cases of toxoplasmosis in humans could be alternatively identified on the basis of the detection of T. gondii DNA in amniotic fluids, tissues, or blood by using the molecular assays such as polymerase chain reaction (PCR) [10,11]. During the last decade, PCR methods have considerable progress for the detection of toxoplasmosis.…”

Section: Introductionmentioning

confidence: 99%

Challenging loop—mediated isothermal amplification (LAMP) technique for molecular detection of Toxoplasma gondii

Fallahi

Mazar

Ghasemian

et al. 2015

Asian Pacific Journal of Tropical Medicine

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Evaluation of a real-time PCR assay based on the single-copy SAG1 gene for the detection of Toxoplasma gondii

Cited by 33 publications

References 13 publications

Partial Protective Effect of Intranasal Immunization with Recombinant Toxoplasma gondii Rhoptry Protein 17 against Toxoplasmosis in Mice

Partial Protective Effect of Intranasal Immunization with Recombinant Toxoplasma gondii Rhoptry Protein 17 against Toxoplasmosis in Mice

Intranasal Immunisation with Recombinant Toxoplasma gondii Actin Partly Protects Mice against Toxoplasmosis

Challenging loop—mediated isothermal amplification (LAMP) technique for molecular detection of Toxoplasma gondii

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