Abstract
A modest amount of cell-free DNA is constantly present in human blood, originating from programmed cell death, apoptosis and rupture of blood cells or pathogens. Acute or chronic cell injury contributes to enhance the pool of circulating nucleic acids, so that their assessment may be regarded as an appealing perspective for diagnosing myocardial ischemia. We performed a search in Medline, Web of Science and Scopus to identify clinical studies that investigated the concentration of cell-free DNA in patients with myocardial ischemia. Overall, eight case-control studies could be detected and reviewed. Although the concentration of cell-free DNA was found to be higher in the diseased than in the healthy population, the scenario was inconclusive due to the fact that the overall number of subjects studied was modest, the populations were unclearly defined, cases and controls were not adequately matched, the methodology for measuring the reference cardiac biomarkers was inadequately described, and the diagnostic performance of cell-free DNA was not benchmarked against highly sensitive troponin immunoassays. Several biological and technical hurdles were also identified in cell-free DNA testing, including the lack of specificity and unsuitable kinetics for early diagnosis of myocardial ischemia, the long turnaround time and low throughput, the need for specialized instrumentation and dedicated personnel, the lack of standardization or harmonization of analytical techniques, the incremental costs and the high vulnerability to preanalytical variables. Hence it seems reasonable to conclude that the analysis of cell-free DNA is not ready for prime time in diagnostics of myocardial ischemia.
References
1. Lippi G, Plebani M. Biomarker research and leading causes of death worldwide: a rather feeble relationship. Clin Chem Lab Med 2013;51:1691–3.10.1515/cclm-2013-0210Search in Google Scholar PubMed
2. Cervellin G, Lippi G. Of MIs and men – a historical perspective on the diagnostics of acute myocardial infarction. Semin Thromb Hemost 2014;40:535–43.10.1055/s-0034-1383544Search in Google Scholar PubMed
3. Thygesen K, Alpert JS, Jaffe AS, Simoons ML, Chaitman BR, White HD; Joint ESC/ACCF/AHA/WHF task force for the universal definition of myocardial infarction. Third universal definition of myocardial infarction. Circulation 2012;126:2020–35.10.1161/CIR.0b013e31826e1058Search in Google Scholar PubMed
4. Thygesen K, Mair J, Giannitsis E, Mueller C, Lindahl B, Blankenberg S, et al; Study group on biomarkers in cardiology of ESC working group on acute cardiac care. How to use high-sensitivity cardiac troponins in acute cardiac care. Eur Heart J 2012;33:2252–7.10.1093/eurheartj/ehs154Search in Google Scholar PubMed
5. Casagranda I, Cavazza M, Clerico A, Galvani M, Ottani F, Zaninotto M, et al. Proposal for the use in emergency departments of cardiac troponins measured with the latest generation methods in patients with suspected acute coronary syndrome without persistent ST-segment elevation. Clin Chem Lab Med 2013;51:1727–37.10.1515/cclm-2013-0423Search in Google Scholar PubMed
6. Agewall S, Giannitsis E, Jernberg T, Katus H. Troponin elevation in coronary vs. non-coronary disease. Eur Heart J 2011;32:404–11.10.1093/eurheartj/ehq456Search in Google Scholar PubMed
7. Zeller T, Ojeda F, Brunner FJ, Peitsmeyer P, Münzel T, Binder H, et al. High-sensitivity cardiac troponin I in the general population – defining reference populations for the determination of the 99th percentile in the Gutenberg Health Study. Clin Chem Lab Med 2014;53:699–706.Search in Google Scholar
8. Lippi G, Cervellin G. Genetic polymorphisms of human cardiac troponins as an unrecognized challenge for diagnosing myocardial injury. Int J Cardiol 2014;171:467–70.10.1016/j.ijcard.2013.12.221Search in Google Scholar PubMed
9. Jung K, Fleischhacker M, Rabien A. Cell-free DNA in the blood as a solid tumor biomarker – a critical appraisal of the literature. Clin Chim Acta 2010;411:1611–24.10.1016/j.cca.2010.07.032Search in Google Scholar PubMed
10. Marzese DM, Hirose H, Hoon DS. Diagnostic and prognostic value of circulating tumor-related DNA in cancer patients. Expert Rev Mol Diagn 2013;13:827–44.10.1586/14737159.2013.845088Search in Google Scholar PubMed
11. Chiu RW, Lo YM. Clinical applications of maternal plasma fetal DNA analysis: translating the fruits of 15 years of research. Clin Chem Lab Med 2013;51:197–204.10.1515/cclm-2012-0601Search in Google Scholar PubMed
12. Capoluongo E, Plebani M. Circulating fetal cell-free DNA and prenatal molecular diagnostics: are we ready for consensus? Clin Chem Lab Med 2014;52:609–11.10.1515/cclm-2014-0066Search in Google Scholar
13. Tong YK, Lo YM. Diagnostic developments involving cell-free (circulating) nucleic acids. Clin Chim Acta 2006;363:187–96.10.1016/j.cccn.2005.05.048Search in Google Scholar
14. Sigdel TK, Sarwal MM. Cell-free DNA as a measure of transplant injury. Clin Transpl 2012:201–5.Search in Google Scholar
15. Danese E, Montagnana M, Fava C, Guidi GC. Circulating nucleic acids and hemostasis: biological basis behind their relationship and technical issues in assessment. Semin Thromb Hemost 2014;40:766–73.10.1055/s-0034-1387923Search in Google Scholar
16. Chen C, Xu J, Huang F. Recent players in the field of acute myocardial infarction biomarkers: circulating cell-free DNA or microRNAs? Int J Cardiol 2013;168:2956–7.10.1016/j.ijcard.2013.03.118Search in Google Scholar
17. Lippi G, Mattiuzzi C, Cervellin G. MicroRNAs for diagnosing myocardial infarction. Advantages and limitations. Int J Cardiol 2013;168:4849–50.10.1016/j.ijcard.2013.07.069Search in Google Scholar
18. Chang CP, Chia RH, Wu TL, Tsao KC, Sun CF, Wu JT. Elevated cell-free serum DNA detected in patients with myocardial infarction. Clin Chim Acta 2003;327:95–101.10.1016/S0009-8981(02)00337-6Search in Google Scholar
19. Ahn SJ, Costa J, Emanuel JR. PicoGreen quantitation of DNA: effective evaluation of samples pre- or post-PCR. Nucleic Acids Res 1996;24:2623–5.10.1093/nar/24.13.2623Search in Google Scholar PubMed PubMed Central
20. Antonatos D, Patsilinakos S, Spanodimos S, Korkonikitas P, Tsigas D. Cell-free DNA levels as a prognostic marker in acute myocardial infarction. Ann N Y Acad Sci 2006;1075:278–81.10.1196/annals.1368.037Search in Google Scholar PubMed
21. Destouni A, Vrettou C, Antonatos D, Chouliaras G, Traeger-Synodinos J, Patsilinakos S, et al. Cell-free DNA levels in acute myocardial infarction patients during hospitalization. Acta Cardiol 2009;64:51–7.10.2143/AC.64.1.2034362Search in Google Scholar PubMed
22. Rainer TH, Lam NY, Man CY, Chiu RW, Woo KS, Lo YM. Plasma beta-globin DNA as a prognostic marker in chest pain patients. Clin Chim Acta 2006;368:110–3.10.1016/j.cca.2005.12.021Search in Google Scholar PubMed
23. Shimony A, Zahger D, Gilutz H, Goldstein H, Orlov G, Merkin M, et al. Cell free DNA detected by a novel method in acute ST-elevation myocardial infarction patients. Acute Card Care 2010;12:109–11.10.3109/17482941.2010.513732Search in Google Scholar PubMed
24. Goldshtein H, Hausmann MJ, Douvdevani A. A rapid direct fluorescent assay for cell-free DNA quantification in biological fluids. Ann Clin Biochem 2009;46:488–94.10.1258/acb.2009.009002Search in Google Scholar PubMed
25. Jing RR, Wang HM, Cui M, Fang MK, Qiu XJ, Wu XH, et al. A sensitive method to quantify human cell-free circulating DNA in blood: relevance to myocardial infarction screening. Clin Biochem 2011;44:1074–9.10.1016/j.clinbiochem.2011.06.083Search in Google Scholar PubMed
26. Cui M, Fan M, Jing R, Wang H, Qin J, Sheng H, et al. Cell-Free circulating DNA: a new biomarker for the acute coronary syndrome. Cardiology 2013;124:76–84.10.1159/000345855Search in Google Scholar PubMed
27. Pagani F, Stefini F, Micca G, Toppino M, Manoni F, Romano L, et al. Multicenter evaluation of the TOSOH AIA-pack second-generation cardiac troponin I assay. Clin Chem 2004;50:1707–9.10.1373/clinchem.2004.035451Search in Google Scholar PubMed
28. Lou X, Hou Y, Liang D, Peng L, Chen H, Ma S, et al. A novel Alu-based real-time PCR method for the quantitative detection of plasma circulating cell-free DNA: sensitivity and specificity for the diagnosis of myocardial infarction. Int J Mol Med 2015;35:72–80.10.3892/ijmm.2014.1991Search in Google Scholar PubMed PubMed Central
29. Apple FS, Ler R, Murakami MM. Determination of 19 cardiac troponin I and T assay 99th percentile values from a common presumably healthy population. Clin Chem 2012;58:1574–81.10.1373/clinchem.2012.192716Search in Google Scholar PubMed
30. Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l’homme. C R Seances Soc Biol Fil 1948;142:241–3.Search in Google Scholar
31. Lippi G, Mattiuzzi C. The biomarker paradigm: between diagnostic efficiency and clinical efficacy. Pol Arch Med Wewn 2015 Feb 10. pii: AOP_15_026. [Epub ahead of print].Search in Google Scholar
32. Lackner KJ. Do we really need high-sensitive troponin immunoassays in the emergency department? Definitely, yes! Clin Chem Lab Med 2014;52:201–4.10.1515/cclm-2013-0604Search in Google Scholar PubMed
33. Lindahl B. Acute coronary syndrome – the present and future role of biomarkers. Clin Chem Lab Med 2013;51:1699–706.10.1515/cclm-2013-0074Search in Google Scholar PubMed
34. Elshimali YI, Khaddour H, Sarkissyan M, Wu Y, Vadgama JV. The clinical utilization of circulating cell free DNA (CCFDNA) in blood of cancer patients. Int J Mol Sci 2013;14:18925–58.10.3390/ijms140918925Search in Google Scholar PubMed PubMed Central
35. Luke JJ, Oxnard GR, Paweletz CP, Camidge DR, Heymach JV, Solit DB, et al; Cell Free DNA Working Group. Realizing the potential of plasma genotyping in an age of genotype-directed therapies. J Natl Cancer Inst 2014;106:dju214.10.1093/jnci/dju214Search in Google Scholar PubMed PubMed Central
36. Plebani M. Harmonization in laboratory medicine: the complete picture. Clin Chem Lab Med 2013;51:741–51.10.1515/cclm-2013-0075Search in Google Scholar PubMed
37. El Messaoudi S, Rolet F, Mouliere F, Thierry AR. Circulating cell free DNA: preanalytical considerations. Clin Chim Acta 2013;424:222–30.10.1016/j.cca.2013.05.022Search in Google Scholar PubMed
38. Lackner KJ. Laboratory diagnostics of myocardial infarction – troponins and beyond. Clin Chem Lab Med 2013;51:83–9.10.1515/cclm-2012-0572Search in Google Scholar PubMed
39. Lippi G, Montagnana M, Aloe R, Cervellin G. Highly sensitive troponin immunoassays: navigating between the scylla and charybdis. Adv Clin Chem 2012;58:1–29.10.1016/B978-0-12-394383-5.00007-2Search in Google Scholar
40. Lippi G, Cervellin G. Choosing troponin immunoassays in a world of limited resources. J Am Coll Cardiol 2013;62:647–8.10.1016/j.jacc.2013.04.022Search in Google Scholar PubMed
41. Turgeon PJ, Sukumar AN, Marsden PA. Epigenetics of cardiovascular disease – a new “beat” in coronary artery disease. Med Epigenet 2014;2:37–52.10.1159/000360766Search in Google Scholar PubMed PubMed Central
42. Lippi G, Mattiuzzi C, Cervellin G. Circulating microRNAs (miRs) for diagnosing acute myocardial infarction: meta-analysis of available studies. Int J Cardiol 2013;167:277–8.10.1016/j.ijcard.2012.09.152Search in Google Scholar PubMed
©2015 by De Gruyter