{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T20:57:07Z","timestamp":1780347427618,"version":"3.54.1"},"reference-count":60,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2016,8,11]],"date-time":"2016-08-11T00:00:00Z","timestamp":1470873600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>The application of molecular technologies to aid diagnosis and management of infectious diseases has had a major impact and many assays are in routine use. Diagnosis of aspergillosis has lagged behind. Lack of standardization and limited commercial interest have meant that PCR was not included in consensus diagnostic criteria for invasive fungal disease. In the last ten years careful evaluation and validation by the Aspergillus European PCR initiative with the development of standardized extraction, amplification and detection protocols for various specimen types, has provided the opportunity for clinical utility to be investigated. PCR has the potential to not only exclude a diagnosis of invasive aspergillosis but in combination with antigen testing may offer an approach for the early diagnosis and treatment of invasive aspergillosis in high-risk populations, with the added benefit of detection of genetic markers associated with antifungal resistance.<\/jats:p>","DOI":"10.3390\/jof2030023","type":"journal-article","created":{"date-parts":[[2016,8,11]],"date-time":"2016-08-11T10:29:29Z","timestamp":1470911369000},"page":"23","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["PCR Technology for Detection of Invasive Aspergillosis"],"prefix":"10.3390","volume":"2","author":[{"given":"Rosemary","family":"Barnes","sequence":"first","affiliation":[{"name":"Department of Medical Microbiology and Infectious Diseases, Cardiff University School of Medicine, Cardiff CF14 4XN, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P.","family":"White","sequence":"additional","affiliation":[{"name":"Public Health Wales Microbiology, Cardiff CF14 4 XW, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2016,8,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"S191","DOI":"10.1080\/13693780600898003","article-title":"Aspergillus PCR\u2014Platforms, strengths and weaknesses","volume":"44","author":"White","year":"2006","journal-title":"Med. Mycol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.idc.2015.10.015","article-title":"Invasive aspergillosis current strategies for diagnosis and management","volume":"30","author":"Cadena","year":"2016","journal-title":"Infect. Dis. Clin. N. Am."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1159\/000196477","article-title":"Pulmonary aspergillosis\u2014Early diagnosis improves survival","volume":"62","author":"Voneiff","year":"1995","journal-title":"Respiration"},{"key":"ref_4","unstructured":"Pasqualotto, A.C. (2010). Aspergillosis: From Diagnosis to Prevention, Springer."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Girmenia, C., Micozzi, A., Gentile, G., Santilli, S., Arleo, E., Cardarelli, L., Capria, S., Minotti, C., Cartoni, C., and Brocchieri, S. (2009). Clinically driven diagnostic antifungal approach in neutropenic patients: A prospective feasibility study. J. Clin. Oncol.","DOI":"10.1200\/JCO.2009.21.8032"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"iii17","DOI":"10.1093\/jac\/dkt391","article-title":"Triggers for driving treatment of at-risk patients with invasive fungal disease","volume":"68","author":"Drgona","year":"2013","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"846","DOI":"10.1038\/bmt.2011.178","article-title":"ECIL recommendations for the use of biological markers for the diagnosis of invasive fungal diseases in leukemic patients and hematopoietic SCT recipients","volume":"47","author":"Marchetti","year":"2012","journal-title":"Bone Marrow Transplant."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1813","DOI":"10.1086\/588660","article-title":"Revised definitions of invasive fungal disease from the European Organization for Research and Treatment of Cancer\/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC\/MSG) Consensus Group","volume":"46","author":"Walsh","year":"2008","journal-title":"Clin. Infect. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Hou, T.Y., Wang, S.H., Liang, S.X., Jiang, W.X., Luo, D.D., and Huang, D.H. (2015). The screening performance of serum 1,3-\u03b2-d-Glucan in patients with invasive fungal diseases: A meta-analysis of prospective cohort studies. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0131602"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Leeflang, M.M.G., Debets-Ossenkopp, Y.J., Wang, J., Visser, C.E., Scholten, R.J.P.M., Hooft, L., Bijlmer, H.A., Reitsma, J.B., Zhang, M., and Bossuyt, P.M. (2015). Galactomannan detection for invasive aspergillosis in immunocompromised patients. Cochrane Database Syst. Rev.","DOI":"10.1002\/14651858.CD007394.pub2"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1007\/s15010-015-0849-8","article-title":"Evaluation of serum 1,3-\u03b2-d-glucan clinical performance: Kinetic assessment, comparison with galactomannan and evaluation of confounding factors","volume":"44","author":"Pini","year":"2016","journal-title":"Infection"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3652","DOI":"10.1128\/JCM.00942-12","article-title":"Diagnostic accuracy of PCR alone compared to galactomannan in bronchoalveolar lavage fluid for diagnosis of invasive pulmonary aspergillosis: A systematic review","volume":"50","author":"Avni","year":"2012","journal-title":"J. Clin. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Zou, M.X., Tang, L.H., Zhao, S.S., Zhao, Z.J., Chen, L.Y., Chen, P., Huang, Z., Li, J., Chen, L., and Fan, X. (2012). Systematic review and meta-analysis of detecting galactomannan in bronchoalveolar lavage fluid for diagnosing invasive aspergillosis. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0043347"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1688","DOI":"10.1086\/647935","article-title":"Bronchoalveolar lavage fluid galactomannan for the diagnosis of invasive pulmonary aspergillosis in patients with hematologic diseases","volume":"49","author":"Maertens","year":"2009","journal-title":"Clin. Infect. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"E1286","DOI":"10.1542\/peds.2007-2117","article-title":"Pediatric invasive aspergillosis: A multicenter retrospective analysis of 139 contemporary cases","volume":"121","author":"Burgos","year":"2008","journal-title":"Pediatrics"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1144","DOI":"10.1093\/cid\/cir122","article-title":"The diagnostic value of halo and reversed halo signs for invasive mold infections in compromised hosts","volume":"52","author":"Georgiadou","year":"2011","journal-title":"Clin. Infect. Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1273","DOI":"10.1086\/657065","article-title":"Probable invasive aspergillosis without prespecified radiologic findings: Proposal for inclusion of a new category of aspergillosis and implications for studying novel therapies","volume":"51","author":"Nucci","year":"2010","journal-title":"Clin. Infect. Dis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1293","DOI":"10.1093\/cid\/civ507","article-title":"Aspergillus polymerase chain reaction: systematic review of evidence for clinical use in comparison with antigen testing","volume":"61","author":"White","year":"2015","journal-title":"Clin. Infect. Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1128\/JCM.02814-15","article-title":"Comparison of performance characteristics of Aspergillus PCR in testing a range of blood-based samples in accordance with international methodological recommendations","volume":"54","author":"Springer","year":"2016","journal-title":"J. Clin. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2832","DOI":"10.1128\/JCM.00905-15","article-title":"Clinical performance of Aspergillus PCR for testing serum and plasma: A study by the European Aspergillus PCR initiative","volume":"53","author":"White","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1231","DOI":"10.1128\/JCM.01767-09","article-title":"Aspergillus PCR: One step closer to standardization","volume":"48","author":"White","year":"2010","journal-title":"J. Clin. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3842","DOI":"10.1128\/JCM.05316-11","article-title":"Evaluation of Aspergillus PCR protocols for testing serum specimens","volume":"49","author":"White","year":"2011","journal-title":"J. Clin. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3753","DOI":"10.1128\/JCM.01466-10","article-title":"Critical stages of extracting DNA from Aspergillus fumigatus in whole-blood specimens","volume":"48","author":"White","year":"2010","journal-title":"J. Clin. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1445","DOI":"10.1128\/JCM.03322-12","article-title":"Multicenter comparison of serum and whole-blood specimens for detection of Aspergillus DNA in high-risk hematological patients","volume":"51","author":"Springer","year":"2013","journal-title":"J. Clin. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2838","DOI":"10.1128\/JCM.00906-15","article-title":"Analytical comparison of in vitro-spiked human serum and plasma for PCR-based detection of Aspergillus fumigatus DNA: A study by the European Aspergillus PCR initiative","volume":"53","author":"Loeffler","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"ref_26","first-page":"1160","article-title":"Comparison of four automated nucleic acid extraction platforms for the recovery of DNA from Aspergillus fumigatus","volume":"63","author":"Perry","year":"2014","journal-title":"J. Clin. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1263","DOI":"10.1093\/cid\/civ555","article-title":"Galactomannan and polymerase chain reaction-based screening for invasive aspergillosis among high-risk hematology patients: A diagnostic meta-analysis","volume":"61","author":"Arvanitis","year":"2015","journal-title":"Clin. Infect. Dis."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2403","DOI":"10.1128\/JCM.00744-13","article-title":"Development and evaluation of a calibrator material for nucleic acid-based assays for diagnosing Aspergillosis","volume":"51","author":"Lyon","year":"2013","journal-title":"J. Clin. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","article-title":"The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments","volume":"55","author":"Bustin","year":"2009","journal-title":"Clin. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Cruciani, M., Mengoli, C., Loeffler, J., Donnelly, P., Barnes, R., Jones, B.L., Klingspor, L., Morton, O., and Maertens, J. (2015). Polymerase chain reaction blood tests for the diagnosis of invasive Aspergillosis in immunocompromised people. Cochrane Database Syst. Rev.","DOI":"10.1002\/14651858.CD009551.pub2"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/S1473-3099(09)70019-2","article-title":"Use of PCR for diagnosis of invasive Aspergillosis: Systematic review and meta-analysis","volume":"9","author":"Mengoli","year":"2009","journal-title":"Lancet Infect. Dis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1016","DOI":"10.1038\/bmt.2010.219","article-title":"Silent and prolonged Aspergillus DNAemia in oncohematological patients receiving antifungal prophylaxis: A new phenomenon with clinical implications","volume":"46","author":"Meije","year":"2011","journal-title":"Bone Marrow Transplant."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2431","DOI":"10.1093\/jac\/dkt227","article-title":"Directed therapy for fungal infections: Focus on Aspergillosis","volume":"68","author":"Barnes","year":"2013","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1510","DOI":"10.1128\/JCM.03189-12","article-title":"Evaluation of Real-Time PCR, Galactomannan Enzyme-Linked Immunosorbent Assay (ELISA), and a Novel lateral-flow device for diagnosis of invasive Aspergillosis","volume":"51","author":"White","year":"2013","journal-title":"J. Clin. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1016\/j.jinf.2013.04.020","article-title":"Prevention and diagnosis of invasive fungal disease in high-risk patients within an integrative care pathway","volume":"67","author":"Barnes","year":"2013","journal-title":"J. Infect."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1093\/cid\/ciu833","article-title":"Serum galactomannan vs. a combination of galactomannan and polymerase chain reaction-based aspergillus dna detection for early therapy of invasive aspergillosis in high-risk hematological patients: A randomized controlled trial","volume":"60","author":"Aguado","year":"2015","journal-title":"Clin. Infect. Dis."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1016\/S1473-3099(13)70076-8","article-title":"Galactomannan and PCR vs. culture and histology for directing use of antifungal treatment for invasive aspergillosis in high-risk haematology patients: A randomised controlled trial","volume":"13","author":"Morrissey","year":"2013","journal-title":"Lancet Infect. Dis."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1111\/bjh.12285","article-title":"Combined real-time PCR and galactomannan surveillance improves diagnosis of invasive Aspergillosis in high risk patients with haematological malignancies","volume":"161","author":"Rogers","year":"2013","journal-title":"Br. J. Haematol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1317","DOI":"10.1016\/j.clinthera.2015.03.021","article-title":"Economic comparison of an empirical vs diagnostic-driven strategy for treating invasive fungal disease in immunocompromised patients","volume":"37","author":"Barnes","year":"2015","journal-title":"Clin. Ther."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Sun, W., Wang, K., Gao, W., Su, X., Qian, Q., Lu, X., Song, Y., Guo, Y., and Shi, Y. (2011). Evaluation of PCR on bronchoalveolar lavage fluid for diagnosis of invasive Aspergillosis: A bivariate metaanalysis and systematic review. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0028467"},{"key":"ref_41","first-page":"89","article-title":"A systematic literature review on the diagnosis of invasive Aspergillosis using polymerase chain reaction (PCR) from bronchoalveolar lavage clinical samples","volume":"24","author":"Tuon","year":"2007","journal-title":"Rev. Iberoam. Micol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"i45","DOI":"10.1093\/jac\/dkq441","article-title":"Optimizing management of invasive mould diseases","volume":"66","author":"Agrawal","year":"2011","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"457","DOI":"10.1007\/s00277-015-2562-1","article-title":"Analysis of the efficiency and costs of antifungal prophylaxis and mycological diagnostics in patients undergoing allogeneic haematopoietic cell transplantation: \u201cReal life\u201d evaluation","volume":"95","author":"Bertz","year":"2016","journal-title":"Ann. Hematol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1696","DOI":"10.1093\/cid\/ciu673","article-title":"Serum galactomannan-based early detection of invasive Aspergillosis in hematology patients receiving effective antimold prophylaxis","volume":"59","author":"Duarte","year":"2014","journal-title":"Clin. Infect. Dis."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1016\/j.cmi.2015.09.009","article-title":"Prospective multicentre PCR-based Aspergillus DNA screening in high-risk patients with and without primary antifungal mould prophylaxis","volume":"22","author":"Springer","year":"2016","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1111\/myc.12322","article-title":"Potential of polymerase chain reaction and galactomannan for the diagnosis of invasive aspergillosis in patients with febrile neutropenia","volume":"58","author":"Aslan","year":"2015","journal-title":"Mycoses"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1093\/mmy\/myu025","article-title":"Performance of MycAssay Aspergillus DNA real-time PCR assay compared with the galactomannan detection assay for the diagnosis of invasive Aspergillosis from serum samples","volume":"52","author":"Danylo","year":"2014","journal-title":"Med. Mycol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Guinea, J., Padilla, C., Escribano, P., Munoz, P., Padilla, B., Gijon, P., and Bouza, E. (2013). Evaluation of MycAssay Aspergillus for diagnosis of invasive pulmonary Aspergillosis in patients without hematological cancer. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0061545"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1139\/w11-064","article-title":"Detection of Aspergillus in lung and other tissue samples using the MycAssay Aspergillus real-time PCR kit","volume":"57","author":"Follett","year":"2011","journal-title":"Can. J. Microbiol."},{"key":"ref_50","first-page":"75","article-title":"Detection of Pneumocystis jirovecii and Aspergillus spp. DNA in bronchoalveolar lavage fluids by commercial real-time PCR assays: Comparison with conventional diagnostic tests","volume":"38","author":"Orsi","year":"2015","journal-title":"New Microbiol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.diagmicrobio.2012.03.001","article-title":"Performance of 2 commercial real-time polymerase chain reaction assays for the detection of Aspergillus and Pneumocystis DNA in bronchoalveolar lavage fluid samples from critical care patients","volume":"73","author":"Orsi","year":"2012","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1111\/myc.12438","article-title":"The effect of clinical characteristics on the performance of galactomannan and PCR for the diagnosis of invasive Aspergillosis in febrile neutropenic patients","volume":"59","author":"Oz","year":"2016","journal-title":"Mycoses"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1007\/s10096-014-2211-y","article-title":"Clinical performance of a commercial real-time PCR assay for Aspergillus DNA detection in serum samples from high-risk patients: Comparison with a galactomannan enzyme immunoassay","volume":"34","author":"Pini","year":"2015","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"4273","DOI":"10.1128\/JCM.05026-11","article-title":"Diagnosis of invasive Aspergillosis by a commercial real-time PCR Assay for Aspergillus DNA in bronchoalveolar lavage fluid samples from high-risk patients compared to a galactomannan enzyme immunoassay","volume":"49","author":"Torelli","year":"2011","journal-title":"J. Clin. Microbiol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2169","DOI":"10.1128\/JCM.00101-11","article-title":"Evaluation of analytical and preliminary clinical performance of myconostica MycAssay Aspergillus When testing serum specimens for diagnosis of invasive Aspergillosis","volume":"49","author":"White","year":"2011","journal-title":"J. Clin. Microbiol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"868","DOI":"10.1128\/JCM.03216-14","article-title":"Validation of a new Aspergillus real-time PCR Assay for direct detection of Aspergillus and azole resistance of Aspergillus fumigatus on bronchoalveolar lavage fluid","volume":"53","author":"Chong","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"2115","DOI":"10.1128\/JCM.00667-15","article-title":"Analytical and clinical evaluation of the pathonostics Aspergenius assay for detection of invasive Aspergillosis and resistance to azole antifungal drugs during testing of serum samples","volume":"53","author":"White","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"O1059","DOI":"10.1111\/1469-0691.12749","article-title":"Polymerase-chain reaction\/electrospray ionization-mass spectrometry for the detection of bacteria and fungi in bronchoalveolar lavage fluids: A prospective observational study","volume":"20","author":"Huttner","year":"2014","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"2283","DOI":"10.1097\/CCM.0000000000001249","article-title":"Rapid diagnosis of infection in the critically ill, a multicenter study of molecular detection in bloodstream infections, pneumonia, and sterile site infections","volume":"43","author":"Vincent","year":"2015","journal-title":"Crit. Care Med."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1007\/s11046-014-9756-2","article-title":"Identification of fungal pathogens by visible microarray system in combination with isothermal gene amplification","volume":"178","author":"Sakai","year":"2014","journal-title":"Mycopathologia"}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/2\/3\/23\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T19:28:12Z","timestamp":1760210892000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/2\/3\/23"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,8,11]]},"references-count":60,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2016,9]]}},"alternative-id":["jof2030023"],"URL":"https:\/\/doi.org\/10.3390\/jof2030023","relation":{},"ISSN":["2309-608X"],"issn-type":[{"value":"2309-608X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,8,11]]}}}