Identification of Circulating miR-22-3p and miR-93-5p as Stable Endogenous Control in Tuberculosis Study
The diagnosis and prognosis of tuberculosis remains challenging and necessitates the development of a new test that can accurately diagnose and monitor treatment responses. In this regard, miRNA is becoming a potential diagnostic and prognostic biomarker which differentiates treatment respondents fr...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-10-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/10/11/868 |
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| author | Workneh Korma Adane Mihret Azeb Tarekegn Yunhee Chang Dasom Hwang Tesfaye Sisay Tessema Hyeyoung Lee |
| author_facet | Workneh Korma Adane Mihret Azeb Tarekegn Yunhee Chang Dasom Hwang Tesfaye Sisay Tessema Hyeyoung Lee |
| author_sort | Workneh Korma |
| collection | DOAJ |
| description | The diagnosis and prognosis of tuberculosis remains challenging and necessitates the development of a new test that can accurately diagnose and monitor treatment responses. In this regard, miRNA is becoming a potential diagnostic and prognostic biomarker which differentiates treatment respondents from non-respondents for various non-infectious and infectious diseases, including tuberculosis. The concentration of miRNAs varies based on cell type, disease, and site of infection, implicating that selection of an optimal reference gene is crucial, and determines the quantification of transcript level and biological interpretation of the data. Thus, the study evaluated the stability and expression level of five candidate miRNAs (let-7i-5p, let-7a-5p, miRNA-16-5p, miRNA-22-3p and miRNA-93-5p), including U6 Small Nuclear RNA (RNU6B) to normalize circulating miRNAs in the plasma of 68 participants (26 healthy controls, 23 latent, and 19 pulmonary tuberculosis infected) recruited from four health centers and three hospitals in Addis Ababa, Ethiopia. The expression levels of miRNAs isolated from plasma of culture confirmed newly diagnosed pulmonary tuberculosis patients were compared with latently infected and non-infected healthy controls. The qPCR data were analyzed using four independent statistical tools: Best Keeper, Genorm, Normfinder and comparative delta-Ct methods, and the data showed that miRNA-22-3p and miRNA-93-5p were suitable plasma reference miRNAs in a tuberculosis study. |
| first_indexed | 2024-03-10T15:22:41Z |
| format | Article |
| id | doaj.art-6bbb0a76b6a3417282af652d098deb22 |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-03-10T15:22:41Z |
| publishDate | 2020-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-6bbb0a76b6a3417282af652d098deb222023-11-20T18:19:32ZengMDPI AGDiagnostics2075-44182020-10-01101186810.3390/diagnostics10110868Identification of Circulating miR-22-3p and miR-93-5p as Stable Endogenous Control in Tuberculosis StudyWorkneh Korma0Adane Mihret1Azeb Tarekegn2Yunhee Chang3Dasom Hwang4Tesfaye Sisay Tessema5Hyeyoung Lee6Molecular diagnostic laboratory, Department of Medical Laboratory Sciences, Yonsei University, Wonju 26493, KoreaArmauer Hansen Research Institute, Addis Ababa 1005, EthiopiaArmauer Hansen Research Institute, Addis Ababa 1005, EthiopiaMolecular diagnostic laboratory, Department of Medical Laboratory Sciences, Yonsei University, Wonju 26493, KoreaMolecular diagnostic laboratory, Department of Medical Laboratory Sciences, Yonsei University, Wonju 26493, KoreaInstitute of Biotechnology, Addis Ababa University, Addis Ababa 1176, EthiopiaMolecular diagnostic laboratory, Department of Medical Laboratory Sciences, Yonsei University, Wonju 26493, KoreaThe diagnosis and prognosis of tuberculosis remains challenging and necessitates the development of a new test that can accurately diagnose and monitor treatment responses. In this regard, miRNA is becoming a potential diagnostic and prognostic biomarker which differentiates treatment respondents from non-respondents for various non-infectious and infectious diseases, including tuberculosis. The concentration of miRNAs varies based on cell type, disease, and site of infection, implicating that selection of an optimal reference gene is crucial, and determines the quantification of transcript level and biological interpretation of the data. Thus, the study evaluated the stability and expression level of five candidate miRNAs (let-7i-5p, let-7a-5p, miRNA-16-5p, miRNA-22-3p and miRNA-93-5p), including U6 Small Nuclear RNA (RNU6B) to normalize circulating miRNAs in the plasma of 68 participants (26 healthy controls, 23 latent, and 19 pulmonary tuberculosis infected) recruited from four health centers and three hospitals in Addis Ababa, Ethiopia. The expression levels of miRNAs isolated from plasma of culture confirmed newly diagnosed pulmonary tuberculosis patients were compared with latently infected and non-infected healthy controls. The qPCR data were analyzed using four independent statistical tools: Best Keeper, Genorm, Normfinder and comparative delta-Ct methods, and the data showed that miRNA-22-3p and miRNA-93-5p were suitable plasma reference miRNAs in a tuberculosis study.https://www.mdpi.com/2075-4418/10/11/868tuberculosismiRNAreference miRNAsqPCR (quantitative polymerase chain reaction)miRNA-22-3pmiRNA-93-5p |
| spellingShingle | Workneh Korma Adane Mihret Azeb Tarekegn Yunhee Chang Dasom Hwang Tesfaye Sisay Tessema Hyeyoung Lee Identification of Circulating miR-22-3p and miR-93-5p as Stable Endogenous Control in Tuberculosis Study Diagnostics tuberculosis miRNA reference miRNAs qPCR (quantitative polymerase chain reaction) miRNA-22-3p miRNA-93-5p |
| title | Identification of Circulating miR-22-3p and miR-93-5p as Stable Endogenous Control in Tuberculosis Study |
| title_full | Identification of Circulating miR-22-3p and miR-93-5p as Stable Endogenous Control in Tuberculosis Study |
| title_fullStr | Identification of Circulating miR-22-3p and miR-93-5p as Stable Endogenous Control in Tuberculosis Study |
| title_full_unstemmed | Identification of Circulating miR-22-3p and miR-93-5p as Stable Endogenous Control in Tuberculosis Study |
| title_short | Identification of Circulating miR-22-3p and miR-93-5p as Stable Endogenous Control in Tuberculosis Study |
| title_sort | identification of circulating mir 22 3p and mir 93 5p as stable endogenous control in tuberculosis study |
| topic | tuberculosis miRNA reference miRNAs qPCR (quantitative polymerase chain reaction) miRNA-22-3p miRNA-93-5p |
| url | https://www.mdpi.com/2075-4418/10/11/868 |
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