Early Detection of Toxoplasma gondii Infection in Mongolian Gerbil by Quantitative Real-Time Pcr

Dai, Fangwei; Zhuo, Xunhui; Kong, Qingming; Du, Jiangtao; Heng, Yu; Zhou, Shasang; Song, Xiaoming; Tong, Qunbo; Lou, Di; Lou, Qi; Lu, Lingqun; Liu, Yu; Xiao-ying, SA; Lu, Shaohong

doi:10.1645/18-95

Cited by 6 publications

(6 citation statements)

References 24 publications

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“…qPCR was performed using a SYBR Premix Ex Taq II (Takara, Tokoyo, Japan) on a Mx3005P real-time PCR System (Stratagene, Waltham, Massachusetts, USA) as described previously. 44 …”

Section: Methodsmentioning

confidence: 99%

“…qPCR was performed using a SYBR Premix Ex Taq II (Takara, Tokoyo, Japan) on a Mx3005P real-time PCR System (Stratagene, Waltham, Massachusetts, USA) as described previously. 44 We established a standard curve to quantify the number of ΔGRA17 tachyzoites from the Ct values obtained from qPCR analysis of tumor tissues. Briefly, DNA was isolated from a 10-fold dilution series of ΔGRA17 tachyzoites (10 6 , 10 5 , 10 4 , 10 3 ).…”

Section: Analysis Of the Immune Status Of The Injected And Distant Tumorsmentioning

confidence: 99%

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Synergy betweenToxoplasma gondiitype I ΔGRA17immunotherapy and PD-L1 checkpoint inhibition triggers the regression of targeted and distal tumors

Zhu

Elsheikha

Wang

et al. 2021

J Immunother Cancer

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BackgroundIn this study, we hypothesize that the ability of the protozoan Toxoplasma gondii to modulate immune response within the tumor might improve the therapeutic effect of immune checkpoint blockade. We examined the synergetic therapeutic activity of attenuated T. gondii RH ΔGRA17 strain and programmed death ligand-1 (PD-L1) treatment on both targeted and distal tumors in mice.MethodsThe effects of administration of T. gondii RH ΔGRA17 strain on the tumor volume and survival rate of mice bearing flank B16-F10, MC38, or LLC tumors were studied. We characterized the effects of ΔGRA17 on tumor biomarkers’ expression, PD-L1 expression, immune cells infiltrating the tumors, and expression of immune-related genes by using immunohistochemistry, immunofluorescence, flow cytometry, NanoString platform, and real-time quantitative PCR, respectively. The role of immune cells in the efficacy of ΔGRA17 plus PD-L1 blockade therapy was determined via depletion of immune cell subtypes.ResultsTreatment with T. gondii ΔGRA17 tachyzoites and anti-PD-L1 therapy significantly extended the survival of mice and suppressed tumor growth in preclinical mouse models of melanoma, Lewis lung carcinoma, and colon adenocarcinoma. Attenuation of the tumor growth was detected in the injected and distant tumors, which was associated with upregulation of innate and adaptive immune pathways. Complete regression of tumors was underpinned by late interferon-gamma-producing CD8+ cytotoxic T cells.ConclusionThe results from these models indicate that intratumoral injection of ΔGRA17 induced a systemic effect, improved mouse immune response, and sensitized immunologically ‘cold’ tumors and rendered them sensitive to immune checkpoint blockade therapy.

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“…qPCR was performed using a SYBR Premix Ex Taq II (Takara, Tokoyo, Japan) on a Mx3005P real-time PCR System (Stratagene, Waltham, Massachusetts, USA) as described previously. 44 …”

Section: Methodsmentioning

confidence: 99%

Section: Analysis Of the Immune Status Of The Injected And Distant Tumorsmentioning

confidence: 99%

Synergy betweenToxoplasma gondiitype I ΔGRA17immunotherapy and PD-L1 checkpoint inhibition triggers the regression of targeted and distal tumors

Zhu

Elsheikha

Wang

et al. 2021

J Immunother Cancer

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“…Traditionally, the quantitative polymerase chain reaction (qPCR) assay has been considered as the gold standard method to detect T. gondii DNA with considerable sensitivity and specificity. , However, qPCR requires thermocycling instrumentation and more than 1 h of time, which limits its field application in remote areas. Isothermal amplification-related techniques for nucleic acid detection can circumvent the limitation of PCR-based assays and have been becoming an alternative to qPCR. , Loop-mediated isothermal amplification (LAMP), one of the most commonly used isothermal techniques, has been developed to detect T. gondii in water, − soil, animals, − and clinical specimens .…”

mentioning

confidence: 99%

RPA/CRISPR/Cas12a-Based On-Site and Rapid Nucleic Acid Detection of Toxoplasma gondii in the Environment

Lei

et al. 2022

ACS Synth. Biol.

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Toxoplasma gondii is an opportunistic pathogen widely distributed within the world, poses a huge threat to human health, and causes significant economic losses to the livestock industry. Herein, we developed a portable one-pot detection of T. gondii by combining recombinase polymerase amplification (RPA) and a clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system. A glass microfiber filter device used for the first step can efficiently extract T. gondii from lowconcentration samples. The lyophilized RPA reagents and Cas12a/crRNA reagents are prestored in one Eppendorf tube, and both reactions can be performed on a low-cost thermal controller (∼37 °C), avoiding the drawbacks of the step-by-step addition of components. The developed RPA/CRISPR/Cas12a system exhibits a high selectivity toward the B1 gene amplicon of T. gondii over other parasites with a limit of detection of 3.3 copies/μL. The visual signal readout can be easily realized by a fluorometer or lateralflow strip. A portable suitcase containing the minimum equipment and lyophilized reagents was adopted for the rapid determination of T. gondii in heavily polluted landfill leachate. This system presents rapidness, robustness and on-site features for the detection of nucleic acids of the parasite, making it a promising tool for field applications in remote areas.

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“…This approach is particularly useful for species that do not have sequence information available. Mongolian gerbil is a gerbil strain that has been widely used in a variety of studies [4,11,28]. In a previous study, we have screened 28 microsatellite marker systems to determine the genetic quality of a closed group of Mongolian gerbils through the use of cross-amplifying technology, using microsatellite sites in mice [6].…”

Section: Discussionmentioning

confidence: 99%

Development of an effective microsatellite marker system to determine the genetic structure of <i>Meriones meridianus</i> populations

Liu

et al. 2020

Exp. Anim.

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Understanding the genetic quality of the gerbil, Meriones meridianus, plays an important role in the study of medical biology. However, no effective system has been established for evaluating a population's genetic diversity to date. In the present study, we established a set of reasonable evaluative systems based on microsatellite markers of the Mongolian gerbil by using the method of cross-amplification of species. Following electrophoresis analysis, short tandem repeat (STR) scanning, and sequencing, 11 microsatellite loci were identified by matching the criteria characteristics and were used to evaluate the genetic diversity of two stocks of Meriones meridianus: Meriones meridianus jei Wang, 1964 (M. m. jei) and Meriones meridianus cryptorhinus Blanford, 1875 (M. m. cryptorhinus) from Xinjiang, China. The microsatellite loci screened were highly polymorphic and were suitable for genetic quality control of Meriones meridianus. In addition, the quality of the non-bred M. m. jei and M. m. cryptorhinus strains in our study is sufficient for them to be promising stocks in the future for the farmed animal industry.

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Early Detection of Toxoplasma gondii Infection in Mongolian Gerbil by Quantitative Real-Time Pcr

Cited by 6 publications

References 24 publications

Synergy betweenToxoplasma gondiitype I ΔGRA17immunotherapy and PD-L1 checkpoint inhibition triggers the regression of targeted and distal tumors

Synergy betweenToxoplasma gondiitype I ΔGRA17immunotherapy and PD-L1 checkpoint inhibition triggers the regression of targeted and distal tumors

RPA/CRISPR/Cas12a-Based On-Site and Rapid Nucleic Acid Detection of Toxoplasma gondii in the Environment

Development of an effective microsatellite marker system to determine the genetic structure of <i>Meriones meridianus</i> populations

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