Improved protocol for plasma microRNA extraction and comparison of commercial kits

We investigated the association between plasma microRNA (miR)-204 and coronary artery calcification (CAC) in patients with type 2 diabetes mellitus (T2DM). We consecutively enrolled 179 individuals with T2DM who underwent coronary computed tomography at Anzhen Hospital from January 2015 to September 2016. The CAC score (CACS) was expressed in Agatston units and >10 Hounsfield units were defined as CAC-positive status. Significant CAC was observed in 98 (54.7%) patients. Plasma miR-204 levels (relative expression) were significantly lower in patients with significant CAC than controls (1.001 ± 0.100 vs 0.634 ± 0.211, P < .001). Plasma miR-204 levels were also negatively correlated with the glycosylated hemoglobin A1c (HbA1c) level (r = −0.702, P < .001), CACS (r = −0.710, P < .001), and the United Kingdom Prospective Diabetes Study (UKPDS) score (r = −0.355, P < .001). After multivariate logistic analyses, plasma miR-204 levels were still significantly and independently associated with the presence of CAC (odds ratio = 0.103, CI = 0.018-0.583, P < .001) after adjustment for conventional risk factors. Receiver operating characteristic curve analysis showed that plasma miR-204 levels can predict the severity and extent of CAC, and the specificity was higher than that of the traditional risk factors UKPDS score and HbA1c. In conclusion, the downregulation of miR-204 was independently associated with CAC in patients with T2DM.

Download Full-text

MPT64 assays for the rapid detection of Mycobacterium tuberculosis

BMC Infectious Diseases ◽

10.1186/s12879-021-06022-w ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Xun-Jie Cao ◽

Ya-Ping Li ◽

Jia-Ying Wang ◽

Jie Zhou ◽

Xu-Guang Guo

Keyword(s):

Mycobacterium Tuberculosis ◽

Rapid Detection ◽

Web Of Science ◽

Mortality Rates ◽

Cochrane Library ◽

Diagnostic Efficiency ◽

Funnel Plot ◽

High Infection ◽

Diagnostic Technology ◽

Commercial Kits

Abstract Background Tuberculosis (TB) is a serious infectious disease caused by Mycobacterium tuberculosis (MTB). An estimated 1.7 billion people worldwide are infected with Mycobacterium tuberculosis (LTBI) during the incubation period without any obvious symptoms. Because of MTB’s high infection and mortality rates, there is an urgent need to develop a fast, portable, and sensitive diagnostic technology for its detection. Methods We included research from PubMed, Cochrane Library, Web of Science, and Embase and extracted the data. MetaDisc and STATA were used to build forest plots, Deek’s funnel plot, Fagan plot, and bivariate boxplot for analysis. Results Forty-six articles were analyzed, the results of which are as follows: sensitivity and specificity were 0.92 (0.91–0.93) and 0.95 (0.94–0.95) respectively. The NLR and PLR were 0.04 (95% CI 0.03–0.07) and 25.32 (95% CI 12.38–51.78) respectively. DOR was 639.60 (243.04–1683.18). The area under the SROC curve (AUC) was 0.99. Conclusions MPT64 exhibits good diagnostic efficiency for MTB. There is no obvious heterogeneity between the three commercial kits.

Download Full-text

Velocity Gradient Separation Reveals a New Extracellular Vesicle Population Enriched in miR-155 and Mitochondrial DNA

Pathogens ◽

10.3390/pathogens10050526 ◽

2021 ◽

Vol 10 (5) ◽

pp. 526

Author(s):

Myriam Vaillancourt ◽

Audrey Hubert ◽

Caroline Subra ◽

Julien Boucher ◽

Wilfried Wenceslas Bazié ◽

...

Keyword(s):

Mitochondrial Dna ◽

Velocity Gradient ◽

Messenger Rna ◽

Sucrose Density Gradient ◽

Extracellular Vesicle ◽

Therapeutic Interventions ◽

Optimal Separation ◽

Velocity Gradients ◽

Commercial Kits ◽

Hiv 1

Extracellular vesicles (EVs) and their contents (proteins, lipids, messenger RNA, microRNA, and DNA) are viewed as intercellular signals, cell-transforming agents, and shelters for viruses that allow both diagnostic and therapeutic interventions. EVs circulating in the blood of individuals infected with human immunodeficiency virus (HIV-1) may provide insights into pathogenesis, inflammation, and disease progression. However, distinguishing plasma membrane EVs from exosomes, exomeres, apoptotic bodies, virions, and contaminating proteins remains challenging. We aimed at comparing sucrose and iodixanol density and velocity gradients along with commercial kits as a means of separating EVs from HIV particles and contaminating protein like calprotectin; and thereby evaluating the suitability of current plasma EVs analysis techniques for identifying new biomarkers of HIV-1 immune activation. Multiple analysis have been performed on HIV-1 infected cell lines, plasma from HIV-1 patients, or plasma from HIV-negative individuals spiked with HIV-1. Commercial kits, the differential centrifugation and density or velocity gradients to precipitate and separate HIV, EVs, and proteins such as calprotectin, have been used. EVs, virions, and contaminating proteins were characterized using Western blot, ELISA, RT-PCR, hydrodynamic size measurement, and enzymatic assay. Conversely to iodixanol density or velocity gradient, protein and virions co-sedimented in the same fractions of the sucrose density gradient than AChE-positive EVs. Iodixanol velocity gradient provided the optimal separation of EVs from viruses and free proteins in culture supernatants and plasma samples from a person living with HIV (PLWH) or a control and revealed a new population of large EVs enriched in microRNA miR-155 and mitochondrial DNA. Although EVs and their contents provide helpful information about several key events in HIV-1 pathogenesis, their purification and extensive characterization by velocity gradient must be investigated thoroughly before further use as biomarkers. By revealing a new population of EVs enriched in miR-155 and mitochondrial DNA, this study paves a way to increase our understanding of HIV-1 pathogenesis.

Download Full-text

Plasma microRNA signature associated with retinopathy in patients with type 2 diabetes

Scientific Reports ◽

10.1038/s41598-021-83047-w ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Donato Santovito ◽

Lisa Toto ◽

Velia De Nardis ◽

Pamela Marcantonio ◽

Rossella D’Aloisio ◽

...

Keyword(s):

Type 2 Diabetes ◽

Diabetic Retinopathy ◽

Vision Loss ◽

External Validation ◽

Diabetic Patients ◽

Disease Biomarkers ◽

Response To Stress ◽

Severity Of The Disease ◽

Plasma Microrna

AbstractDiabetic retinopathy (DR) is a leading cause of vision loss and disability. Effective management of DR depends on prompt treatment and would benefit from biomarkers for screening and pre-symptomatic detection of retinopathy in diabetic patients. MicroRNAs (miRNAs) are post-transcriptional regulators of gene expression which are released in the bloodstream and may serve as biomarkers. Little is known on circulating miRNAs in patients with type 2 diabetes (T2DM) and DR. Here we show that DR is associated with higher circulating miR-25-3p (P = 0.004) and miR-320b (P = 0.011) and lower levels of miR-495-3p (P < 0.001) in a cohort of patients with T2DM with DR (n = 20), compared with diabetic subjects without DR (n = 10) and healthy individuals (n = 10). These associations persisted significant after adjustment for age, gender, and HbA1c. The circulating levels of these miRNAs correlated with severity of the disease and their concomitant evaluation showed high accuracy for identifying DR (AUROC = 0.93; P < 0.001). Gene ontology analysis of validated targets revealed enrichment in pathways such as regulation of metabolic process (P = 1.5 × 10–20), of cell response to stress (P = 1.9 × 10–14), and development of blood vessels (P = 2.7 × 10–14). Pending external validation, we anticipate that these miRNAs may serve as putative disease biomarkers and highlight novel molecular targets for improving care of patients with diabetic retinopathy.

Download Full-text

Evaluation of three commercial kits for the confirmation of antibodies to human immunodeficiency virus (HIV-1).

Clinical Chemistry ◽

10.1093/clinchem/34.9.1930 ◽

1988 ◽

Vol 34 (9) ◽

pp. 1930-1930 ◽

Cited By ~ 1

Author(s):

S D Pearson ◽

C T Wittwer ◽

K O Ash

Keyword(s):

Human Immunodeficiency Virus ◽

Immunodeficiency Virus ◽

Commercial Kits ◽

Hiv 1

Download Full-text

Identification of plasma microRNA-21 as a biomarker for early detection and chemosensitivity of non-small cell lung cancer

Chinese Journal of Cancer ◽

10.5732/cjc.010.10522 ◽

2011 ◽

Vol 30 (6) ◽

pp. 407-414 ◽

Cited By ~ 147

Author(s):

Juan Wei ◽

Wen Gao ◽

Cheng-Jun Zhu ◽

Yi-Qian Liu ◽

Zhu Mei ◽

...

Keyword(s):

Lung Cancer ◽

Early Detection ◽

Small Cell Lung Cancer ◽

Cell Lung Cancer ◽

Small Cell ◽

Small Cell Lung ◽

Plasma Microrna

Download Full-text

The triiodothyronine uptake test: an assessment of methods.

Clinical Chemistry ◽

10.1093/clinchem/27.7.1272 ◽

1981 ◽

Vol 27 (7) ◽

pp. 1272-1276 ◽

Cited By ~ 5

Author(s):

L R Witherspoon ◽

S E Shuler ◽

M M Garcia

Keyword(s):

Functional Status ◽

Coefficient Of Variation ◽

Temperature Effects ◽

Thyroxine Binding Globulin ◽

Wide Range ◽

Free Thyroxine Index ◽

Total Thyroxine ◽

Multiple Measurements ◽

Uptake Test ◽

Commercial Kits

Abstract How well the free thyroxine index reflects thyroid functional status depends on the degree to which the triiodothyronine uptake test normalizes the effects of thyroxine binding protein concentrations on the total thyroxine concentration. We examined eight triiodothyronine uptake tests in which were used different secondary binders representative of those available in commercial kits. The relation between triiodothyronine uptake and thyroxine-binding globulin concentrations was established by use of sera from euthyroid individuals. We examined the effects of both high (greater than 20 mg/L) and low (less than 10 mg/L) thyroxine-binding globulin concentrations on triiodothyronine uptake. The precision of each assay, expressed as within- and between-run coefficient of variation, was calculated from multiple measurements on high, low, and midrange triiodothyronine uptake serum pools. The effects of variation in temperature and in exposure times were examined. The clinical most useful assays exhibited the ability to reflect a wide range of thyroxine-binding globulin concentrations and demonstrated little or no time or temperature effects.

Download Full-text

MicroRNA-451 as an Early Predictor of Chronic Kidney Disease in Diabetic Nephropathy

International Journal of Nephrology ◽

10.1155/2020/8075376 ◽

2020 ◽

Vol 2020 ◽

pp. 1-7

Author(s):

Mostafa Abdelsalam ◽

A. M. Wahab ◽

Maysaa El Sayed Zaki ◽

Mohamad Motawea

Keyword(s):

Chronic Kidney Disease ◽

Diabetic Nephropathy ◽

Kidney Disease ◽

Early Diagnosis ◽

Serum Creatinine ◽

High Sensitivity ◽

Urinary Albumin ◽

Significant Difference ◽

Plasma Microrna ◽

Stage Renal Disease

Background. Diabetes mellitus is the leading cause of end-stage renal disease worldwide. Microalbuminuria is the cornerstone for the diagnosis of diabetic nephropathy. However, it is an inadequate marker for early diagnosis. MicroRNAs are not only new and promising markers for early diagnosis but also, but they may also play a role in the prevention of disease progression. Methods. This study included ninety patients with type 2 DM in addition to 30 control subjects. MicroRNA-451 expression in blood and plasma using real-time PCR was evaluated in addition to the classic diabetic nephropathy markers (serum creatinine, urinary albumin, and eGFR). Results. There was a significant difference between the studied groups versus control regarding serum creatinine, eGFR, urinary, and plasma microRNA-451 with p=0.0001. Patients with eGFR 60 ml/min/1.73 m2 showed a significantly higher plasma microRNA-451 (29.6 ± 1.6) and significantly lower urinary microRNA-451 (21 ± 0.9) in comparison to patients with eGFR >60 ml/min/1.73 m2 and p=0.0001. eGFR showed a positive correlation with urinary microRNA-451 and negative correlation with both plasma microRNA-451 and urinary albumin. Both plasma and urinary microRNA-451 are highly sensitive and specific markers for chronicity in diabetic nephropathy patients with sensitivity of 90.9% and 95.5% and specificity of 67.6% and 95.6%, respectively. Conclusion. MicroRNA-451 is a promising early biomarker for chronic kidney disease in diabetic nephropathy with high sensitivity and specificity.

Download Full-text