title: Evaluation of a multiplex ligation-dependent probe amplification assay for the detection of respiratory pathogens in oncological patients.
abstract: BACKGROUND
  Respiratory tract infections are widespread and may cause significant morbidity and mortality in immunosuppressed populations such as oncological patients.
  
  
  OBJECTIVE
  The RealAccurate Respiratory RT PCR Kit covering 14 respiratory viruses was compared to the RespiFinder Smart22, a broad-spectrum multiplex ligation-dependent probe amplification (MLPA) test, targeting 22 viral and bacterial respiratory pathogens.
  
  
  METHODS
  After verification of its analytical performance, the clinical performance of the RespiFinder Smart22 was evaluated by re-analysis of 96 respiratory samples from oncological patients. Additionally, the time to result (TTR) of both methods was compared.
  
  
  RESULTS
  The analytical performance of the RespiFinder Smart22 fulfilled all previously specified criteria. Concordant results in both assays were achieved in 74.0% of all clinical specimens and in 91.2% when only positive results were taken into account. The RespiFinder Smart22 yielded additional results in a total of 22 (22.9% of 96) samples due to higher test sensitivity and a broader, highly multiplexed spectrum of pathogens. The TTR of a typical routine test consisting of three samples were 206 and 356 min for the RealAccurate Respiratory RT PCR Kit and the RespiFinder Smart22, respectively. However, hands-on time was reduced by 59.0% applying the MLPA method.
  
  
  CONCLUSIONS
  In our hands, the RespiFinder Smart22 showed excellent analytical performance while hands-on time was halved in comparison to the RT PCR method. Regarding the clinical evaluation, the MLPA method provided additional results in 22.9% (22/96) of specimens due to its comprehensive format, higher test sensitivity and the capability to detect 22 pathogens compared to 14 with the RealAccurate Respiratory RT PCR Kit.
authors.affiliation: analyse BioLab, Elisabethinen Hospital Linz, Eisenhandstrasse 4-6, 4020 Linz, Austria.
authors.forename: Lucia
authors.initials: L
authors.lastname: Berning
authors.affiliation: Department of Virology, Medical University of Vienna, Kinderspitalgasse 15, 1095 Vienna, Austria.
authors.forename: Stephan W
authors.initials: SW
authors.lastname: Aberle
authors.affiliation: Department of Virology, Medical University of Vienna, Kinderspitalgasse 15, 1095 Vienna, Austria.
authors.forename: Benedikt
authors.initials: B
authors.lastname: Simon
authors.affiliation: analyse BioLab, Elisabethinen Hospital Linz, Eisenhandstrasse 4-6, 4020 Linz, Austria.
authors.forename: Christoph
authors.initials: C
authors.lastname: Luger
authors.affiliation: analyse BioLab, Elisabethinen Hospital Linz, Eisenhandstrasse 4-6, 4020 Linz, Austria.
authors.forename: Petra
authors.initials: P
authors.lastname: Apfalter
authors.affiliation: 1st Internal Department - Hematology with Stem Cell Transplantation, Hemostaseology and Medical Oncology, Elisabethinen Hospital Linz, Fadingerstrasse 1, 4020 Linz, Austria.
authors.forename: Sigrid
authors.initials: S
authors.lastname: Machherndl-Spandl
authors.affiliation: analyse BioLab, Elisabethinen Hospital Linz, Eisenhandstrasse 4-6, 4020 Linz, Austria. Electronic address: heidrun.kerschner@analyse.eu.
authors.forename: Heidrun
authors.initials: H
authors.lastname: Kerschner
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001424:Bacterial Infections / Q000175:diagnosis* / Q000382:microbiology; D005260:Female; D006801:Humans; D016867:Immunocompromised Host; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009369:Neoplasms / Q000150:complications*; D012141:Respiratory Tract Infections / Q000175:diagnosis*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D013997:Time Factors; D014777:Virus Diseases / Q000175:diagnosis* / Q000821:virology; D014780:Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D055815:Young Adult
keywords: Hands on time; MLPA; Multiplex detection; Oncology; Respiratory tract infection; Time to result