Evaluation of Enzyme Immunoassay Method versus Real-time PCR for Diagnosis of Rotavirus Infection in Acute Infantile Diarrhea

Background: Acute infectious gastroenteritis is a common cause of fatality between children in the developing countries which is usually due to viral etiology. Rotavirus is a ds-RNA (60-80nm), non-enveloped virus with a segmented genome. Group (A) of the virus is an important human pathogen that accounts for (90%) of the isolates. An easy, rapid, non-expensive and sensitive method is needed to detect this virus for clinical controlling. The objective of this study is to evaluate Enzyme immunoassay technique versus Quantitative real-time PCR in the diagnosis of infection with Rotavirus in the children with acute diarrhea. Methodology: This study was conducted on (75) infants and young children, from The Pediatric Department at Tanta University Hospitals in the period from December 2019 to March 2020 and were diagnosed according to history and clinical examination using Vesikari scoring system for acute severe gastroenteritis. Also, 10 healthy infants and children were taken as a control group. Stool samples were obtained from the patients and the controls. These specimens were tested with ELISA and Quantitative real-time PCR for detection of Rotavirus in stool. Results: The study revealed that 62 patients (82.6 %) were positive by ELISA and 74 cases (98.6 %) were positive with real time RT-PCR. Additionally, all the control group gave negative results by the two techniques. Conclusion: Enzyme immunoassay is an accurate and suitable method as a routine diagnostic measure for Rotavirus that can run a large number of samples. But, it is expensive when used for a single sample. Quantitative real time PCR was more sensitive and specific measure that can detect Rotavirus RNA in too minimal amounts in stools.

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Evaluation of Galactomannan Enzyme Immunoassay and Quantitative Real-Time PCR for the Diagnosis of Invasive Pulmonary Aspergillosis in a Rat Model

Journal of Microbiology and Biotechnology ◽

10.4014/jmb.1401.01067 ◽

2014 ◽

Vol 24 (8) ◽

pp. 1044-1050 ◽

Cited By ~ 6

Author(s):

Jian-cong Lin ◽

Yan-li Xing ◽

Wen-ming Xu ◽

Ming Li ◽

Pang Bo ◽

...

Keyword(s):

Enzyme Immunoassay ◽

Real Time ◽

Rat Model ◽

Real Time Pcr ◽

Invasive Pulmonary Aspergillosis ◽

Quantitative Real Time Pcr ◽

Pulmonary Aspergillosis

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Galactomannan enzyme immunoassay and quantitative Real Time PCR as tools to evaluate the exposure and response in a rat model of aspergillosis after posaconazole prophylaxis

Enfermedades Infecciosas y Microbiología Clínica ◽

10.1016/j.eimc.2015.11.008 ◽

2016 ◽

Vol 34 (9) ◽

pp. 571-576 ◽

Cited By ~ 2

Author(s):

Emilio Cendejas-Bueno ◽

Agustina Forastiero ◽

Isabel Ruiz ◽

Emilia Mellado ◽

María José Buitrago ◽

...

Keyword(s):

Enzyme Immunoassay ◽

Real Time ◽

Rat Model ◽

Real Time Pcr ◽

Quantitative Real Time Pcr

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Recruitment of cyanobacteria by reverse transcription quantitative real-time PCR based on expression of Microcystis gene

PeerJ ◽

10.7717/peerj.7188 ◽

2019 ◽

Vol 7 ◽

pp. e7188

Author(s):

Long Yu ◽

Xiaofei Wu ◽

Yang Yu ◽

Limei Shi ◽

Min Zhang

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Lake Taihu ◽

Rna Extraction ◽

Sybr Green ◽

Quantitative Detection ◽

Control Group ◽

Quantitative Real Time Pcr ◽

Total Rna ◽

Pcr Method

In this study, a SYBR Green quantitative real-time PCR method was established and applied. Relative expression of the synthetic genes from Microcystis gas vesicles (gvpC), algal toxin genes (mcyA), and polysaccharides (espL) from water and sediments of Meiliang Bay and from the center of Lake Taihu were tested from January to June, 2017. Indoor Microcystis aeruginosa was used as the control group. The kit for total RNA extraction in Microcystis was optimized. Results showed that the optimized kit extracted high-concentrations and high-quality total RNA from Microcystis. The extraction purity and concentration were significantly higher than those extracted by the original kit. The transcription level of gvpC increased gradually until a peak was reached in March. However, expression of gvpC decreased continuously at the proliferating and floating stages of Cyanobacterial biomass. The maximum level of expression of gvpC in April in comparison to expression of mcyA in March occurred first. We found that the SYBR Green qRT-PCR method, which is characterized by high specificity, repeatability, is rapid, and can be used for quantitative detection of expression of gvpC, mcyA, and espL. The recruitment of cyanobacteria is the process in which cyanobacteria in the sediment began to regain their activity, started to grow and migrated to the water column.

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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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Detection of Streptococcus pneumoniae in Sputum Samples by Real- Time PCR.

Anti-Infective Agents ◽

10.2174/2211352518999200629165108 ◽

2020 ◽

Vol 18 ◽

Author(s):

Pegah Shakib ◽

Mohammad Reza Zolfaghari

Keyword(s):

Streptococcus Pneumoniae ◽

Real Time ◽

Real Time Pcr ◽

False Negative ◽

Cross Sectional Study ◽

Laboratory Culture ◽

Cross Sectional ◽

Negative Results ◽

False Negative Results ◽

Pcr Method

Background: Conventional laboratory culture-based methods for diagnosis of Streptococcus pneumoniae are time-consuming and yield false negative results. Molecular methods including real-time (RT)-PCR rapid methods and conventional PCR due to higher sensitivity and accuracy have been replaced instead traditional culture assay. The aim of the current study was to evaluate lytA gene for detection of Streptococcus pneumoniae in the cerebrospinal fluid of human patients with meningitis using real-time PCR assay. Material and Methods: In this cross-sectional study, a total of 30 clinical specimens were collected from patients in a period from September to December 2018. In order to evaluate the presence of lytA gene, conventional and real-time PCR methods were used without culture. Results: From 30 sputum samples five (16.66%) isolates were identified as S. pneumoniae by lytA PCR and sequencing. Discussion: In this research, an accurate and rapid real-time PCR method was used, which is based on lytA gene for diagnosis of bacteria so that it can be diagnosed. Based on the sequencing results, the sensitivity for detection of lytA gene was 100% (5/5).

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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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