title: Effectiveness of PCR and Immunofluorescence Techniques for Detecting Human Cytomegalovirus in Blood and Bronchoalveolar Lavage Fluid.
abstract: Current diagnostic methods allow a rapid and reliable detection of active human cytomegalovirus (hCMV) infection by identifying the presence of pp65 CMV antigen or CMV DNA in peripheral blood and affected organs. The goal of this study was to evaluate the effectiveness of CMV detection in blood and organ-specific biological material, such as bronchoalveolar lavage fluid (BALF), by comparing two standard diagnostic methods, immunofluorescence (IF) and the real-time polymerase chain reaction (PCR). We evaluated 25 patients with concomitant respiratory disease who were referred to our hospital for diagnosis due to suspected acute CMV infection. The presence of hCMV was concomitantly evaluated by IF and PCR in 16 peripheral blood samples. In two patients, we observed positive results for both IF and PCR, and in two other patients the results were discordant. Of 11 patients, CMV DNA was detected in six BALF samples, and in one blood plasma sample. Real-time PCR detected CMV DNA in 54.6 % of BALF samples and 12.0 % of blood samples, while indirect IF testing confirmed antigenemia in 12.5 % of blood samples. The results from our study suggest that the IF method is as effective as PCR for detecting an ongoing CMV infection in blood samples. However, real-time PCR was much more effective at detecting CMV DNA in BALF compared to blood samples. Our results suggest that the biological material being tested during CMV diagnosis should be derived directly from the virally infected organ(s).
authors.affiliation: Department of Genetics and Clinical Immunology, National Institute of Tuberculosis and Lung Diseases, 26 Płocka St, 01-138, Warsaw, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Roży
authors.affiliation: Department of Genetics and Clinical Immunology, National Institute of Tuberculosis and Lung Diseases, 26 Płocka St, 01-138, Warsaw, Poland.
authors.forename: K
authors.initials: K
authors.lastname: Duk
authors.affiliation: Department of Genetics and Clinical Immunology, National Institute of Tuberculosis and Lung Diseases, 26 Płocka St, 01-138, Warsaw, Poland.
authors.forename: B
authors.initials: B
authors.lastname: Szumna
authors.affiliation: Department of Genetics and Clinical Immunology, National Institute of Tuberculosis and Lung Diseases, 26 Płocka St, 01-138, Warsaw, Poland.
authors.forename: P
authors.initials: P
authors.lastname: Skrońska
authors.affiliation: Third Department of Lung Disease, National Institute of Tuberculosis and Lung Diseases, Warsaw, Poland.
authors.forename: D
authors.initials: D
authors.lastname: Gawryluk
authors.affiliation: Department of Genetics and Clinical Immunology, National Institute of Tuberculosis and Lung Diseases, 26 Płocka St, 01-138, Warsaw, Poland. j.chorostowska@igichp.edu.pl.
authors.forename: J
authors.initials: J
authors.lastname: Chorostowska-Wynimko
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001992:Bronchoalveolar Lavage Fluid / Q000821:virology*; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism / Q000821:virology; D004279:DNA, Viral / Q000235:genetics*; D005260:Female; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D010750:Phosphoproteins / Q000276:immunology / Q000378:metabolism*; D012372:ROC Curve; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D019562:Viral Load; D014763:Viral Matrix Proteins / Q000276:immunology / Q000378:metabolism*; D055815:Young Adult
chemical_list: D004279:DNA, Viral; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa
keywords: Blood; Bronchoalveolar lavage fluid; Human cytomegalovirus; Lungs; Viral infection; pp65 antigen