First molecular detection of Toxoplasma gondii in vegetable samples in China using qualitative, quantitative real-time PCR and multilocus genotyping

AbstractToxoplasma gondii infection is becoming increasing problem in China but there is no data concerning contamination of vegetables intended for consumption with this parasite. The aim of the present study was to investigate fresh vegetables originated from open markets located in the Xining City, the Qinghai-Tibet Plateau (QTP), P.R. China for their contamination with T. gondii. A total of 279 fresh vegetable samples were collected and analysed using real-time PCR assay targeting B1 gene and multilocus genotyping. T. gondii DNA was found in 10 (3.6%) samples tested; eight of them represented T. gondii type I and remaining two T. gondii type II. The approximate level of contamination of positive vegetables samples, estimated based on quantitative real-time PCR (qPCR), ranged between less than one and 27000 T. gondii oocysts per sample, with majority not exceeding several oocysts per sample. The results of the study confirmed that T. gondii is present in vegetables offered in open markets in the Qinghai province, P.R. China; eating them unwashed and raw may therefore pose a threat to consumers. This is the first investigation describing T. gondii detection in fresh vegetables intended for consumption collected from the territory of P.R. China using sensitive molecular tools.

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Toxoplasma gondii and pre-treatment protocols for polymerase chain reaction analysis of milk samples: a field trial in sheep from Southern Italy

Italian Journal of Food Safety ◽

10.4081/ijfs.2017.6501 ◽

2017 ◽

Vol 6 (1) ◽

Cited By ~ 1

Author(s):

Alice Vismarra ◽

Elena Barilli ◽

Maura Miceli ◽

Carlo Mangia ◽

Cristina Bacci ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Real Time ◽

Dna Extraction ◽

Real Time Pcr ◽

Southern Italy ◽

Raw Milk ◽

Type I ◽

Treatment Protocols ◽

Milk Samples ◽

Pre Treatment

Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii. Ingestion of raw milk has been suggested as a risk for transmission to humans. Here the authors evaluated pre-treatment protocols for DNA extraction on T. gondii tachyzoite-spiked sheep milk with the aim of identifying the method that resulted in the most rapid and reliable PCR positivity. This protocol was then used to analyze milk samples form sheep from three different farms in southern Italy, including Real Time PCR for DNA quantification and PCR-RFLP for genotyping. The pre-treatment protocol using EDTA and Tris-HCl to remove casein gave the best results in the least amount of time compared to the others on spiked milk samples. One sample of 21 collected from sheep farms was positive on one-step PCR, Real Time PCR and resulted in a Type I genotype at one locus (SAG3). Milk usually contains a low number of tachyzoites and this could be a limiting factor for molecular identification. Our preliminary data has evaluated a rapid, cost-effective and sensitive protocol to treat milk before DNA extraction. The results of the present study also confirm the possibility of T. gondii transmission through consumption of raw milk and its unpasteurized derivatives.

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

Journal of Parasitology ◽

10.1645/18-95 ◽

2019 ◽

Vol 105 (1) ◽

pp. 52 ◽

Cited By ~ 1

Author(s):

Fangwei Dai ◽

Xunhui Zhuo ◽

Qingming Kong ◽

Jiangtao Du ◽

Haijie Yu ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Early Detection ◽

Real Time ◽

Mongolian Gerbil ◽

Real Time Pcr ◽

Quantitative Real Time Pcr ◽

Toxoplasma Gondii Infection

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The first detection ofToxoplasma gondiiDNA in environmental air samples using gelatine filters, real-time PCR and loop-mediated isothermal (LAMP) assays: qualitative and quantitative analysis

Parasitology ◽

10.1017/s0031182017001172 ◽

2017 ◽

Vol 144 (13) ◽

pp. 1791-1801 ◽

Cited By ~ 5

Author(s):

ANNA LASS ◽

BEATA SZOSTAKOWSKA ◽

KRZYSZTOF KORZENIEWSKI ◽

PANAGIOTIS KARANIS

Keyword(s):

Toxoplasma Gondii ◽

Real Time ◽

Real Time Pcr ◽

Microscopic Analysis ◽

Detection Methods ◽

Type I ◽

Air Samples ◽

Two Samples ◽

Environmental Air ◽

Tract Infections

SUMMARYToxoplasma gondiiinfections are acquired through the ingestion of oocysts present in the environment. However, there is no data about their occurrence in the air or about airborne transmission of these infections. In the present paper, we report on the identification ofT. gondiiusing rapid molecular detection methods, supported by microscopic analysis, in environmental air samples. A total of 71 samples were collected, using gelatine filters, from kitchen gardens, recreational areas and sandpits located in northern and north-eastern Poland. Material recovered from the filters was analysed using real-time PCR and loop-mediated isothermal assays targeting theT. gondiiB1 gene.Toxoplasma gondiiDNA was found in two samples, as confirmed by both molecular assays. Genotyping at the SAG2 locus showedToxoplasmaSAG2 type I. Moreover, the presence ofT. gondiioocysts was confirmed in one of the positive samples with the use of microscopy. The results showed thatT. gondiimay be present in environmental air samples and that respiratory tract infections may play a role in the high prevalence of toxoplasmosis in humans and animals. To the best of our knowledge, this is the first epidemiological evidence that oro-fecal and foodborne toxoplasmosis may be traceable to an airborne respiratory origin and that this may represent a new, previously unknown transmission route for this disease.

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Detection of Toxoplasma gondii in amniotic fluid using quantitative real-time PCR method

Orvosi Hetilap ◽

10.1556/oh.2007.27971 ◽

2007 ◽

Vol 148 (20) ◽

pp. 935-938 ◽

Cited By ~ 1

Author(s):

Bálint Nagy ◽

Levente Lázár ◽

Gyula Richárd Nagy ◽

Zoltán Bán ◽

Zoltán Papp

Keyword(s):

Toxoplasma Gondii ◽

Amniotic Fluid ◽

Real Time ◽

Real Time Pcr ◽

Quantitative Real Time Pcr ◽

Pcr Method

Bevezetés: A Toxoplasma gondii (T. gondii) által okozott fertőzés általában tünetmentes, vagy csak enyhe panaszokat okoz. A terhesekre azonban veszélyt jelent a parazita. A T. gondiiv al fertőzött anyáról a kórokozó transplacentalisan a magzatot is megfertőzheti és congenitalis toxoplasmosist okozhat, amely súlyos magzati elváltozásokat idézhet elő. A korai diagnózis a kezelés sikerét növeli. A congenitalis toxoplasmosis kimutatható szerológiai és molekuláris biológiai módszerekkel. Cél: A T. gondii kimutatása a kvantitatív valós idejű PCR-módszerrel magzatvízből. Módszerek: A magzatvízből a DNS-t szilika adszorbciós módszerrel izoláltuk. A T. gondii kimutatása 74 mintából történt meg kvantitatív valós idejű PCR-módszerrel. Eredmények: A szerzők 74 magzatvízmintából hat esetben mutatták ki a parazita jelenlétét. Konklúzió: A kvantitatív valós idejű PCR-módszer megnöveli a kimutatás gyorsaságát és érzékenységét, valamint lehetőséget nyújt a kórokozó mennyiségi meghatározására is. Ez új utat nyithat a prognózis megállapítására, valamint a kezelés eredményességének a monitorizálására.

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

Asian Pacific Journal of Tropical Medicine ◽

10.4103/1995-7645.250341 ◽

2019 ◽

Vol 12 (1) ◽

pp. 26 ◽

Cited By ~ 1

Author(s):

Ehsan Ahmadpour ◽

Elmira Zargami ◽

Mahmoud Mahami-Oskouei ◽

Adel Spotin ◽

Abbas Shahbazi ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Pregnant Women ◽

Real Time ◽

Real Time Pcr ◽

Quantitative Real Time Pcr ◽

Toxoplasma Gondii Infection

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Validation of Reference Genes for Gene Expression Studies in Virus-Infected Nicotiana benthamiana Using Quantitative Real-Time PCR

PLoS ONE ◽

10.1371/journal.pone.0046451 ◽

2012 ◽

Vol 7 (9) ◽

pp. e46451 ◽

Cited By ~ 176

Author(s):

Deshui Liu ◽

Lindan Shi ◽

Chenggui Han ◽

Jialin Yu ◽

Dawei Li ◽

...

Keyword(s):

Gene Expression ◽

Real Time ◽

Real Time Pcr ◽

Nicotiana Benthamiana ◽

Reference Genes ◽

Quantitative Real Time Pcr ◽

Expression Studies ◽

Gene Expression Studies

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KLK11 promotes the activation of mTOR and protein synthesis to facilitate cardiac hypertrophy

BMC Cardiovascular Disorders ◽

10.1186/s12872-021-02053-y ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Yi Wang ◽

Hongjuan Liao ◽

Yueheng Wang ◽

Jinlin Zhou ◽

Feng Wang ◽

...

Keyword(s):

Protein Synthesis ◽

Angiotensin Ii ◽

Cardiac Hypertrophy ◽

Western Blot ◽

Real Time ◽

Real Time Pcr ◽

Mtor Signaling ◽

Heart Weight ◽

Cardiomyocyte Hypertrophy ◽

Quantitative Real Time Pcr

Abstract Background Cardiovascular diseases have become the leading cause of death worldwide, and cardiac hypertrophy is the core mechanism underlying cardiac defect and heart failure. However, the underlying mechanisms of cardiac hypertrophy are not fully understood. Here we investigated the roles of Kallikrein 11 (KLK11) in cardiac hypertrophy. Methods Human and mouse hypertrophic heart tissues were used to determine the expression of KLK11 with quantitative real-time PCR and western blot. Mouse cardiac hypertrophy was induced by transverse aortic constriction (TAC), and cardiomyocyte hypertrophy was induced by angiotensin II. Cardiac function was analyzed by echocardiography. The signaling pathway was analyzed by western blot. Protein synthesis was monitored by the incorporation of [3H]-leucine. Gene expression was analyzed by quantitative real-time PCR. Results The mRNA and protein levels of KLK11 were upregulated in human hypertrophic hearts. We also induced cardiac hypertrophy in mice and observed the upregulation of KLK11 in hypertrophic hearts. Our in vitro experiments demonstrated that KLK11 overexpression promoted whereas KLK11 knockdown repressed cardiomyocytes hypertrophy induced by angiotensin II, as evidenced by cardiomyocyte size and the expression of hypertrophy-related fetal genes. Besides, we knocked down KLK11 expression in mouse hearts with adeno-associated virus 9. Knockdown of KLK11 in mouse hearts inhibited TAC-induced decline in fraction shortening and ejection fraction, reduced the increase in heart weight, cardiomyocyte size, and expression of hypertrophic fetal genes. We also observed that KLK11 promoted protein synthesis, the key feature of cardiomyocyte hypertrophy, by regulating the pivotal machines S6K1 and 4EBP1. Mechanism study demonstrated that KLK11 promoted the activation of AKT-mTOR signaling to promote S6K1 and 4EBP1 pathway and protein synthesis. Repression of mTOR with rapamycin blocked the effects of KLK11 on S6K1 and 4EBP1 as well as protein synthesis. Besides, rapamycin treatment blocked the roles of KLK11 in the regulation of cardiomyocyte hypertrophy. Conclusions Our findings demonstrated that KLK11 promoted cardiomyocyte hypertrophy by activating AKT-mTOR signaling to promote protein synthesis.

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