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

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Asian Pacific Journal of Tropical Medicine 2015年第5期8卷 366-372页

作者： Shirzad Fallahi Zahra Arab Mazar Mehrdad Ghasemian Ali Haghighi Department of Medical Parasitology and Mycology School of MedicineLorestan University of Medical Sciences Department of Medical Parasitology and Mycology School of MedicineShahid Beheshti University of Medical Sciences Department of Medical Parasitology and Mycology School of MedicineIran University of Medical Sciences

Objective:To compare analytical sensitivity and specificity of a newly described DNA amplification technique.LAMP and nested PCR assay targeting the RE and Bl genes for the detection of Toxoplasma gondii(T.gondii) DNA.Methods:The analytical sensitivity of LAMP and ncstcd-PCR was obtained against 10-fold serial dilutions of T.gondii DNA ranging from 1 ng to 0.01 fg.DNA samples of other parasites and human chromosomal DNA were used to determine the specificity of molecular assays.Results:After testing LAMP and nesled-PCR in duplicate,the detection limit of RE-LAMP.B1-LAMP,RE-nested PCR and B1-nested PCR assays was one fg.100 fg,1 pg and 10 pg of T.gondii DNA respectively.All the LAMP assays and nested PCRs were 100% specific.The RE-LAMP assay revealed the most sensitivity for the detection of T.gondii DNA.Conclusions:The obtained results demonstrate that the LAMP technique has a greater sensitivity for detection of T.gondii.Furthermore,these findings indicate that primers based on the RE are more suitable than those based on the B1 gene.However,the B1-LAMP assay has potential as a diagnostic tool for detection of T.gondii.

关键词： LAMP Nested-PCR Molecular detection Toxoplasma gondii

Influence of Toxoplasma gondii on in vitro proliferation and apoptosis of hepatoma carcinoma H7402 cell

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Asian Pacific Journal of Tropical Medicine 2016年第1期9卷 60-63页

作者： Gang Wang Ming Gao Department of Emergency People's Hospital of Zhengzhou Department of Oncology the First Affiliated Hospital of Zhengzhou University

Objective:To discuss the influence of tachyzoite of Toxoplasma gondii（T.gondii） RH strain on proliferation and apoptosis of hepatoma carcinoma（HCC） H7402 cell.Methods:The HCC H7402 cell in logarithmic phase and tachyzoite of T.gondii RH strain in different concentrations（1×107/mL,2×107/mL.4×107/mL,8×107mL and 16×107/mL） were co-cultured.CCK-8was utilized to determine the inhibition rate of T.gondii tachyzoite on H7402 cell growth.Flow cytometry was used to detect the change of cell cycle.RT-PCR method was used to detect the expression of cyclinB1 and cdc2-two genes related to cell cycle.Western blot method was used to detect the expression of apoptosis-related proteins Caspase-3 and Bcl-2.Results:The tachyzoite of T.gondii RH strain can inhibit the proliferation of HCC H7402 cells.The inhibition rate of tumor cell growth increased with the increase of concentration of T.gondii tachyzoite.With the increase of concentration of T.gondii tachyzoite,the proportion of G0/G1 phase of H7402 cell increased,the proportion of S phase decreased,and PI value decreased accordingly.The expression of cyclinB1 and cdc2 genes decreased with the increase of the concentration of T.gondii tachyzoite.With the increase of the concentration of tachyzoite of T.gondii RH strain,the expression quantity of Caspase-3 in H7402 cell increased,but the expression quantity of Bcl-2protein decreased.Conclusions:T.gondii can inhibit the in vitro proliferation of HCC H7402 cell,and induce its apoptosis.This effect shows a trend of concentration-dependent increase.Moreover,it is related to the down-regulation of cyclinB1 and cdc2（cell cycle-related genes）,the increase of apoptosis-related protein Caspase-3.and the decreasc of Bcl-2 expression.

关键词： Toxoplasma gondii Hepatoma carcinoma H7402 cell line Cell proliferation

Diagnosis of Toxoplasma gondii infection in pregnant women using automated chemiluminescence and quantitative real time PCR

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Asian Pacific Journal of Tropical Medicine 2019年第1期12卷 26-31页

作者： Ehsan Ahmadpour Elmira Zargami Mahmoud Mahami-Oskouei Adel Spotin Abbas Shahbazi Hossein Samadi Kafil Saba Rajabi Paria Alizadeh Yagoob Azadi Reza Bahaj Firouz Shahrivar Aleksandra Barac Infectious and Tropical Disease Research Center Tabriz University of Medical Sciences Students Research Committee Tabriz University of Medical Sciences Department of Parasitology and Mycology Faculty of Medicine Tabriz University of Medical Sciences Drug Applied Research Center Tabriz University of Medical Sciences Clinic for Infectious and Tropical Diseases Clinical Centre of Serbia Faculty of Medicine University of Belgrade

Objective: To identify serodiagnosis and quantification of Toxoplasma gondii(T. gondii) infection among pregnant women in Salmas, northwest of Iran. Methods: In this crosssectional study, 276 blood samples were collected from pregnant women referred to the health care centers in Salmas city. The demographic variables were also recorded. Titers of antiToxoplasma IgM and IgG antibodies(Ab) were determined using the chemiluminescence immunoassay. Quantitative real-time PCR targeting the T. gondii repeated element gene was also performed on the blood sample. Results: Out of all, 19.92%(55/276) and 2.17%(6/276) patients were seropositive for anti-Toxoplasma IgG and IgM Ab, respectively. Moreover, the presence of T. gondii DNA was observed in 12.31%(34/276) blood samples. A significant relationship was observed between the IgG Ab seropositivity and contact with the cat as a risk factor(P=0.022). Conclusions: The seroprevalence rate of T. gondii infection in pregnant women is relatively low. Consequently, the seronegative pregnant women are at risk, and a considerable rate of positive blood samples for the presence of parasite's DNA should not be ignored. Besides, quantitative real-time PCR could be considered as an accurate method for diagnosis of acute toxoplasmosis especially when the precise results are of the most importance in pregnancy. Limiting contact with cats is also suggested for pregnant women.

关键词： Toxoplasmosis Toxoplasma gondii Chemiluminescence immunoassay Real time PCR Pregnancy

Seroprevalence of Toxoplasma gondii infection in dogs and cats in Zhenjiang City,Eastern China

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Asian Pacific Journal of Tropical Biomedicine 2014年第9期4卷 725-728页

作者： Qing-Xin Liu Shuai Wang Li-Qun Wang Jun Xing Wen-Jue Gao Guo-Fang Liu Bin Zhao Hai-Bin Zhang Li-Hua Gao Department of Animal Husbandry and Veterinary Medicine.Jiangsu Polytechnic College of Agriculture and Forestry College of Veterinary Medicine Nanjing Agricultural University

Objective:To determine the seroprevalence of Toxoplasma gondii(T.gondii) infection in dogs and cats in Zhenjiang City,Jiangsu Province.Eastern China,and to evaluate the main associated risk factors relating to exposure to 71 gondii in this region.Methods:Sera from 160 clogs and 116 cats from Zhenjiang City were tested for anti-T.gondii antibodies using EUSA.The seropositivity by area of activity,sex and age was analyzed.Results:Overall.21 dogs(13.l%) and 24 cats(20.7%) had antibodies to T.gondii.The infection rate in stray dogs(38.7%) and cats(28.6%! was significantly higher(P0.05).A high proportion of dogs at 3 to 6 years of age were positive to T.gondii(20.0%)while cats with relatively high seropositivity rates were at 0 to 1 year of age(33.3%).Conclusions:The prevalence of T.gondii infection in dogs and cats in Zhenjiang City was high,which is probably the main source of T.gondii infection in this area.

关键词： Toxoplasma gondii Dog Cat Seroprevalence Zhenjiang China

Toxoplasma gondii Infection in Risk Population Groups from Andes Peruvian

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Open Journal of Medical Microbiology 2020年第1期10卷 26-32页

作者： Jhosef Quispe Pari Ramirez Yauyo Almidon Aquino Mallqui Adauto Salome Ochoa Armida Rojas Raul Montalvo Faculty of Human Medicine “Universidad Nacional del Centro del Perú” Huancayo Peru “Hospital Nacional Ramiro Prialé Prialé” Huancayo Peru

Toxoplasmosis is a parasitic infection whose clinical presentations depend on the human’s immune response. They can range from asymptomatic to rapidly progressive and deadly. There are no data on this disease in our population. Therefore, we have proposed to know the seroprevalence of toxoplasmosis in population risk groups through a descriptive, exploratory study in the 3 most populated districts of the province of Huancayo. The samples were processed by ELISA method and there were a total of 169 samples, 85 pregnant women and 84 cancer patients. The seroprevalence found in pregnant women was 44.7% and in cancer patients it was 26.2%. Finally, the only risk factor associated with positive serology was the place of origin, having a higher risk for those living in the district of Huancayo and Chilca compared to El Tambo (p < 0.05).

关键词： Toxoplasmosis Seroprevalence Toxoplasma gondii Antibodies IgG

Analysis Concentration of Toxoplasma gondiion Anti-Toxoplasma IgG-IgM Antibody Levels, and the Outcomes of Pregnancy in Mice Balb/c

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Open Journal of Obstetrics and Gynecology 2017年第3期7卷 281-289页

作者： Tigor Peniel Simanjuntak Mochammad Hatta Robert H. Sirait Marni Br Karo Lenny Irmawaty Sirait Tetty Rina Aritonang Syahrul Rauf Ressy Dwiyanti Department of Obstetrics and Gynecology Faculty of Medicine Christian University of Indonesia Jakarta Indonesia Molecular Biology and Immunology Laboratory Faculty of Medicine Hasanuddin University Makassar Indonesia Department of Anaestesiology Faculty of Medicine Christian University of Indonesia Jakarta Indonesia Midwife Program of Medistra Health Higher School Jakarta Indonesia Department of Obstetrics and Gynecology Faculty of Medicine Hasanuddin University Makassar Indonesia Department of Medical Microbiology Faculty of Medicine Tadulako University Palu Indonesia

Introduction: Toxoplasma gondii is an obligate intracellular protozoan parasite that can infect any warm blood vertebrae, and if first trimester pregnant woman infected, it may cause abortion. The objective is to prove the effect of the Toxoplasma gondii concentration in anti-toxoplasma IgG-IgM antibody levels, and the outcomes of Balb/c mice pregnancies. Materials and Methods: The study was conducted in Balb/c mice with inclusion criteria, and was conditioned pregnant. The pathogen strains of Toxoplasma gondii tachyzoite injected intraperitoneally. The blood samples were taken serially to be tested for anti-toxoplasma IgG-IgM antibody levels. After the mice were injected with tachyzoite, they are assessed every day to observe their body weight, vaginal bleeding, and labor. Anti-toxoplasma IgG-IgM antibody levels examined using qualitative mouse IgG-IgM antibody ELISA KIT. Results: Anti-toxoplasma IgM antibody levels increased significantly after 24 hours of injection tachyzoites in all dose groups, and remained high through day 21. Anti-toxoplasma antibody IgG levels increased significantly after 72 hours post injection and remained elevated until day 21. The incidence of abortion is 100% in mice which injected tachyzoite levels 1 × 103 and 1 × 104, and the incidence of abortion approximately 2 - 4 days post injection. 100% of mice that were injected with tachyzoites 1 × 101 and 1 × 102 have labor at term. Physical anomaly was found in baby mice from mice that were injected with tachyzoite 1 × 102. Conclusion: There is a significant correlation between the concentrations of Toxoplasma gondii tachyzoite with anti-toxoplasma IgG-IgM antibody levels, and there is a significant relationship between the concentrations of tachyzoite with abortion.

关键词： Toxoplasma gondii Tachyzoite Anti-Toxoplasma IgG-IgM Antibody Levels Pregnancy Abortion Labor

Infection of Toxoplasma gondiiin Humans and Livestock Animals: An Emerging Silent Threat for Bangladesh

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Open Journal of Medical Microbiology 2018年第4期8卷 109-117页

作者： Taibur Rahman Atiqur Rahman Sajib Chakraborty Department of Biochemistry and Molecular Biology University of Dhaka Dhaka Bangladesh

Toxoplasma gondii is an intracellular, zoonotic protozoan parasite that causes toxoplasmosis. It can potentially infect almost all mammalian and avian hosts including one-third of the human population world-wide. The major target group of the parasite includes immunocompromised patients (e.g. AIDS, cancer, organ transplantation) and fetus bearing pregnant women where it develops toxoplasmic encephalitis, myocarditis, chorioretinitis and abnormal fetal brain development or stillbirths respectively. In this review, we have presented the current status of T. gondii infection in livestock animals and human population in Bangladesh to assess the country-wide relative risk. Although exact prevalence is difficult to predict due to the scarcity of data, nevertheless existing literature suggests that 16% - 39% humans and 8% - 70% domestic animals are infected with T. gondii, which implies Bangladeshi population is at high risk of toxoplasmosis. Furthermore, we have proposed a potential area of research to decipher the genetic diversity and transmission routes of T. gondii infection into Bangladeshi population.

关键词： Toxoplasma gondii Seroprevalence Livestock Animals Humans Bangladesh

CD4 T-Lymphocytes Count in HIV-Toxoplasma gondiiCo-Infected Pregnant Women Undergoing a Prevention of Mother-to-Child Transmission Program

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Journal of Biosciences and Medicines 2018年第4期6卷 76-84页

作者： Gnatoulma Katawa Malewe Kolou Liza Koboyo Nadjir Essoham Ataba Gatigbene Bomboma Simplice Damintoti Karou Ecole Superieure des Techniques Biologiques et Alimentaires (ESTBA-UL) Universite de Lome Lomé Togo Laboratoire de Biologie Moleculaire et d’Immunologie (BIOLIM) Universite de Lome Lomé Togo

Toxoplasma gondii (T. gondii) is a parasite responsible of toxoplasmosis, a disease often asymptomatic but with serious consequences in pregnant women and immunocompromised subjects. Objective: This study aimed to investigate the impact of T. gondii infection on CD4+ T lymphocytes count in HIV-infected pregnant women. Methods: This was a cross-sectional study of pregnant women co-infected by HIV and T. gondii. The study was conducted from January to July 2016 at the Prevention of Mother-to-Child Transmission of HIV (PMTCT) sites in the Health District of Lacs in Togo. Diagnosis of HIV was performed by immuno-chromatographic methods with Determine TM HIV-1/2 and immuno-filtration with Tri-Dot HIV-1 and 2 kits. Presence of anti-toxoplasmic IgG and IgM antibodies was established via enzyme immunoassay using ELISA-BIOREX®kit. Flow cytometry was used to count CD4+ T lymphocytes. Results: Our study found that of the 4599 pregnant women, 111 (2.41%) were HIV-positive. Among them, 109 (98.20%) were infected by HIV-1 and 2 (1.98%) by HIV-2. Antibodies against T. gondii were detected in 5.36% (IgM), 25% (IgG) and 3.57% (both IgM and IgG) of HIV 56 infected women. There was no significant difference between CD4 cell count in HIV (+)/T. gondii IgM (-)/IgG (-) infected pregnant women (378.8 ± 222.8 cell//μl) compared to HIV (+)/T. gondii/IgM (+) (457.3 ± 183.3 cell//μl), HIV (+)/T. gondii IgG (+) (419.4 ± 287.3 cell//μl) and HIV (+)/T. gondii IgM/IgG (+) (480.5 ± 252.4 cell/μl). Conclusion: This study showed that intracellular parasite T. gondii did not alter CD4+ T lymphocytes count in HIV/T. gondii co-infected pregnant women.

关键词： HIV Toxoplasma gondii Co-Infection CD4+ T Lymphocytes Count

Psychiatric Variables in Cytomegalovirusor Toxoplasma gondii—Infected Pregnants: Prospects for Cure with a Program of “Counseling about Behavioral Change”

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Open Journal of Preventive Medicine 2015年第7期5卷 315-320页

作者： Concetta De Pasquale Maria Anna Coniglio Maria Luisa Pistorio Nunzio Massimo Scalisi Federica Scalia Carmela Maria Costanzo Marina Marranzano Guido Scalia Department of Medical Surgical Sciences and Advanced Technologies “G.F. Ingrassia” University of Catania Catania Italy Vascular Surgery and Organ Transplant Unit Department of Surgery Transplantation and Advanced technologies University Hospital of Catania Catania Italy Clinical Virology Unit Department of Biomedical Sciences University Hospital of Catania Catania Italy

TORCH complex, one of the potential infections that may occur during pregnancy, may contribute also to prenatal pregnant women’s anxiety and depression. The aim of this study was to explore the presence of psychiatric symptoms, in relation with infection of TORCH agents, specifically the Toxoplasma gondii (T. gondii) and Cytomegalovirus (CMV). The study was conducted on 58 pregnant women recruited from the Clinical Virology Unit, A.O.U. Policlinico-Vittorio Emanuele, P.O. “Gaspare Rodolico”, from September 2012 to March 2014. Psychiatric symptoms were evaluated through the Symptom Checklist-90 Revised (SCL-90 R) in pregnant women with CMV or T. gondii infection. Moreover, pregnant women were invited to participate in a program of “Counseling about behavioral change”, an effective psychotherapeutic training for the development of new motivational strategies to the infection acceptance, the greater self-confidence and greater adherence to treatment. The age of the subjects was positively correlated with Depression (r = 0.119, p ? 0.178, p ? 0.231, r = ? 0.320, p ? 0.208, p ? 0.105, p ? 0.236, p < 0.05). Finally, we can notice an improvement of the variables Anxiety and Somatization in the whole sample after the attendance at the counseling psychotherapic program. Our study suggests the presence of anxiety and somatization in pregnant women with infection, expression of a phase of emotional fragility with the concern of transmitting a serious disease in the unborn child. The program “Counseling about behavioral change” allowed us to create an atmosphere of trust and to understand the usefulness of “empathic listening”. Giving correct and accurate information about the disease reassures the patient and prevents the realization of negative thoughts and further concern and discomfort.

关键词： Pregnancy Psychiatric Symptoms Somatization Toxoplasma gondii Cytomegalovirus