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

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

Scientific Reports ◽

10.1038/s41598-019-54073-6 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 4

Author(s):

Anna Lass ◽

Liqing Ma ◽

Ioannis Kontogeorgos ◽

Xueyong Zhang ◽

Xiuping Li ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Real Time ◽

Real Time Pcr ◽

Tibet Plateau ◽

Type I ◽

Quantitative Real Time Pcr ◽

Vegetable Samples ◽

Multilocus Genotyping ◽

Fresh Vegetables ◽

Open Markets

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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Prospective Study of Diagnosis of Toxoplasma gondii Infection by Elisa and Real-time PCR in Immunocompromised Patients

Frontiers in Immunology ◽

10.3389/conf.fimmu.2014.04.00004 ◽

2014 ◽

Vol 5 ◽

Author(s):

BERREDJEM Hajira ◽

BECHEKER Imène ◽

BENLAIFA Meriem ◽

BENSAID Maya Nadine ◽

BERREDJEM Karima ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Prospective Study ◽

Real Time ◽

Real Time Pcr ◽

Immunocompromised Patients ◽

Toxoplasma Gondii Infection

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Evaluation of significant bacteriuria in pregnant women using quantitative real-time PCR

Journal of obstetrics and women s diseases ◽

10.17816/jowd65450-56 ◽

2016 ◽

Vol 65 (4) ◽

pp. 50-56

Author(s):

Tatyana A. Khusnutdinova ◽

Elena V Shipitsina ◽

Yulia A. Savochkina ◽

Olga Yu. Timoshina ◽

Elena V. Rybina ◽

...

Keyword(s):

Pregnant Women ◽

Real Time ◽

Sensitivity And Specificity ◽

Real Time Pcr ◽

Urine Culture ◽

Urine Samples ◽

Quantitative Real Time Pcr ◽

Diagnostic Characteristics ◽

Significant Bacteriuria ◽

Tract Infections

Introduction. Urinary tract infections are the most common infections in obstetrics and gynecology. Bacteriological method to investigate urine is laborious and time-consuming, therefore development of accurate and rapid methods for the detection of significant bacteriuria is important. Objective. Evaluation of quantitative real-time PCR based approach for the detection of significant bacteriuria in pregnant women. Material and methods. A retrospective investigation of mid-stream urine samples obtained from pregnant women was performed. Urine culture was performed using quantitative method, and a case was considered as significant bacteriuria if ≥ 105 CFU/ml were detected. Urine samples were analyzed for main uropathogens / groups of uropathogens using quantitative multiplex real-time PCR. Diagnostic characteristics of PCR were computed relative to the results of urine culture. Results. In total, 896 urine samples were tested. Of them, significant bacteriuria was found in 28 cases (3%). The frequency of detection of Escherichia coli was 50%, Enterococcus spp. — 25%, Klebsiella spp. — 7%, Proteus spp. and S. saprophyticus 4% each, Streptococcus spp. — 14%. Sensitivity and specificity of the detection of significant bacteriuria using quantitative real-time PCR for the majority of bacterial species / groups were 99% to 100%. Sensitivity and specificity of the quantitative real-time PCR based method were 96% and 98%, respectively. Conclusions. Prevalence of significant bacteriuria among pregnant women is 3%. Half of the uropathogens isolated from pregnant women with bacteriuria are E. coli. Sensitivity and specificity of quantitative PCR for the detection of significant bacteriuria are 96% and 98%, respectively.

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Diagnosis of Toxoplasmosis in Pregnant Women Using Enzyme Immunoassay and Nested Real Time PCR in Al-Najaf city/Iraq

International Journal of Pharmaceutical Research ◽

10.31838/ijpr/2020.12.02.0022 ◽

2020 ◽

Vol 12 (02) ◽

Keyword(s):

Pregnant Women ◽

Enzyme Immunoassay ◽

Real Time ◽

Real Time Pcr

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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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