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Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR
Published on: January 22, 2014
Differential expression of circulating microRNAs in serum: Potential biomarkers to track Japanese encephalitis virus
Manjari Baluni1, Sneha Ghildiyal1, Tanzeem Fatima1
1Department of Microbiology, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, Uttar Pradesh, India.
Abstract:
Japanese encephalitis is one of the serious vector-borne viral encephalitis diseases found worldwide and poses a major threat to public health. Most Japanese encephalitis virus (JEV) infections are subclinical; only 1: 250 to 1:1000 infected persons develop clinical presentations. Delay in proper diagnosis of JE affects the timeliness of treatment initiation and increases the mortality rate in patients. Therefore, there is an extreme need to develop potential biomarkers, which might improve the diagnosis and can become the basis for development of new therapeutics. The microRNAs (miRNAs/or miRs) are small noncoding RNAs of 17-24 nucleotides that are known to regulate about 60% of human genes. Although miRNAs have been found to regulate various aspects of innate and adaptive immune responses, less information on circulating miRNAs in JE is known. The study of JEV infected human serum miRNAs will provide novel information for the diagnosis of JE as well as for the improvement of disease outcome. Total RNA, including miRNA, was extracted from serum followed by the complementary DNA (cDNA) synthesis by using sequence-specific primers. cDNA was amplified using target-specific TaqMan MicroRNA Assay. Real-time polymerase chain reaction data was normalized using both exogenous (cel-miR-39) and endogenous (hsa-miR-93) controls. We have found significantly altered expression of miR-155 and miR-21 in serum of JEV infected patients as compared to healthy controls, revealing their role as a a noninvasive biomarker in JE. A significant correlation between miRNAs and JE was observed that offers the basis for miRNAs to serve as a new component to develop possible therapeutic strategies for JE in near future.
Insights
Japanese encephalitis (JE) is a serious viral disease. Researchers identified miR-155 and miR-21 in patient serum as potential noninvasive biomarkers for improved JE diagnosis and future therapeutic strategies.
Area of Science:
- * Neuroscience and Virology
- * Molecular Biology and Immunology
Background:
- * Japanese encephalitis (JE) is a significant global public health concern, caused by the Japanese encephalitis virus (JEV).
- * Clinical manifestations of JE occur in a small fraction of infected individuals, highlighting the need for early and accurate diagnosis.
- * Current diagnostic delays in JE can lead to increased mortality, emphasizing the need for novel diagnostic biomarkers.
Purpose of the Study:
- * To investigate the role of circulating microRNAs (miRNAs) as potential noninvasive biomarkers for Japanese encephalitis.
- * To identify specific miRNAs in the serum of JEV-infected patients that correlate with disease presence.
- * To explore the potential of these miRNAs for improving JE diagnosis and guiding therapeutic development.
Main Methods:
- * Extraction of total RNA, including miRNA, from the serum of JEV-infected patients and healthy controls.
- * Synthesis of complementary DNA (cDNA) using sequence-specific primers.
- * Quantification of miRNA expression via real-time polymerase chain reaction (PCR) using TaqMan assays and normalization with control miRNAs.
Main Results:
- * Significantly altered expression levels of miR-155 and miR-21 were detected in the serum of JEV-infected patients compared to healthy individuals.
- * These specific miRNAs demonstrated a significant correlation with Japanese encephalitis.
- * The findings suggest a potential role for miR-155 and miR-21 as noninvasive biomarkers for JE.
Conclusions:
- * Circulating miR-155 and miR-21 show promise as noninvasive biomarkers for the diagnosis of Japanese encephalitis.
- * The identified miRNAs offer a basis for developing new diagnostic tools and therapeutic strategies for JE.
- * Further research into miRNA-mediated mechanisms in JE could lead to improved patient outcomes.
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