<clinical_study>
  <!-- This xml conforms to an XML Schema at:
    https://clinicaltrials.gov/ct2/html/images/info/public.xsd -->
  <required_header>
    <download_date>ClinicalTrials.gov processed this data on September 20, 2023</download_date>
    <link_text>Link to the current ClinicalTrials.gov record.</link_text>
    <url>https://clinicaltrials.gov/ct2/show/NCT01695512</url>
  </required_header>
  <id_info>
    <org_study_id>University of Heidelberg</org_study_id>
    <secondary_id>Grant Funding number</secondary_id>
    <nct_id>NCT01695512</nct_id>
  </id_info>
  <brief_title>Diagnostic Study of Combined Biomnarker Testing in Bronchoalveolar Lavage Samples of Immunocompromised Patients</brief_title>
  <official_title>Open, Prospective, Multicenter Trial to Evaluate the Clinical Significance of Combined Serological (Galactomannan ELISA, 1-&gt;3-β-D Glucan Assay) and Molecular (Nested Aspergillus PCR Assay, Multifungal DNA Microarray) Diagnostic Assays to Detect and Characterize Fungal Pathogens in Bronchoalveolar Lavage (BAL) and Blood Samples of Hematological High Risk Patients and to Detect Point Mutations Conferring Azole Resistance</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Heidelberg University</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>Gilead Sciences</agency>
      <agency_class>Industry</agency_class>
    </collaborator>
  </sponsors>
  <source>Heidelberg University</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
  </oversight_info>
  <brief_summary>
    <textblock>
      The aim of our prospective and multicentre diagnostic study is therefore to elucidate on the&#xD;
      sensitivity and specificity rates of these serologic markers in combination with molecular&#xD;
      tools (both an Aspergillus specific and a multifungal PCR based assay), as serologic mark-ers&#xD;
      are not pathogen-specific, and furthermore to define species-specific cut-off values for BDG&#xD;
      in BAL samples.&#xD;
&#xD;
      Additionally, if genomic material of Aspergillus fumigatus is detected by PCR in a clinical&#xD;
      sample, we investigate fungal DNA for point mutations in the cyp51A gene mediating&#xD;
      resis-tance against common mould-active triazoles with novel rapid, sensitive and specific,&#xD;
      non-culture-based PCR-assays and sequencing to optimize antifungal treatment as early as&#xD;
      pos-sible.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      The major problem in managing life threatening invasive fungal infections in patients (pts)&#xD;
      with acute leukemia and pts after allogeneic hematopoietic stem cell transplantation is the&#xD;
      lack of sensitive and specific diagnostic tools to identify fungal pathogens reliably and&#xD;
      early in the course of the disease. Because of deep neutropenia and low platelet count,&#xD;
      invasive diagnostic procedures are rarely feasible in time, therefore bronchoscopy with BAL&#xD;
      is the method of choice in diagnosing pulmonary infections, as the sensitivity of&#xD;
      culture-based methods from blood is low, especially in mould infections. Indirect methods,&#xD;
      so-called surrogate markers, are becoming increasingly important in this clinical setting.&#xD;
      These serologic markers, mainly galactomannan (GM) and recently 1(1,3)-β-D-glucan (BDG), have&#xD;
      been validated in clinical trials for blood samples, however the clinical significance of&#xD;
      testing BAL samples is up to now only based on retrospective data for GM and has not been&#xD;
      reported yet for BDG.&#xD;
&#xD;
      The aim of our prospective and multicentre diagnostic study is therefore to elucidate on the&#xD;
      sensitivity and specificity rates of these serologic markers in combination with molecular&#xD;
      tools (both an Aspergillus specific and a multifungal PCR based assay), as serologic markers&#xD;
      are not pathogen-specific, and furthermore to define species-specific cut-off values for BDG&#xD;
      in BAL samples.&#xD;
&#xD;
      Additionally, if genomic material of Aspergillus fumigatus is detected by PCR in a clinical&#xD;
      sample, we investigate fungal DNA for point mutations in the cyp51A gene mediating resistance&#xD;
      against common mould-active triazoles with novel rapid, sensitive and specific,&#xD;
      non-culture-based PCR-assays and sequencing to optimize antifungal treatment as early as&#xD;
      possible.&#xD;
&#xD;
      This study aims to improve the early, sensitive and specific diagnosis of invasive pulmonary&#xD;
      fungal infections and detect azole resistance patterns, it might impact on the prognosis in&#xD;
      hematologic patients; therefore antifungal treatment data and clinical outcome will be&#xD;
      recorded.&#xD;
&#xD;
      Clinical samples (BAL and blood) from approximately 100 pts suffering from acute leukemia and&#xD;
      pts after allogeneic stem cell transplantation with febrile neutropenia and lung infiltrates&#xD;
      diagnosed in a chest CT scan suggestive for fungal infection will be investigated after pts's&#xD;
      informed consent in a multicentre, prospective trial. Pts will undergo standardized&#xD;
      diagnostic imaging and microbiological procedures for identification of the underlying&#xD;
      infectious pathogen. BAL and blood samples will be tested additionally for GM, BDG and with a&#xD;
      diagnostic nested Aspergillus PCR assay, a multifungal DNA-Microarray and an azole resistance&#xD;
      PCR assay. The molecular assays were established by our group and encompass the detection of&#xD;
      Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, Candida albicans, Candida&#xD;
      dubliniensis, Candida glabrata, Candida lusitaniae, Candida tropicalis, Fusarium oxysporum,&#xD;
      Fusarium solani, Mucor racemosus, Rhizopus microsporus, Scedosporium prolificans,&#xD;
      Trichosporon asahii and identify the three most common point mutations that confer resistance&#xD;
      to triazoles like voriconazole or posaconazole.&#xD;
&#xD;
      Results of other diagnostic means including culture findings as well as patients' clinical&#xD;
      data (e.g. duration of neutropenia, underlying disease and outcome including follow-up data&#xD;
      concerning antifungal treatment and mortality attributable to fungal infections) will be&#xD;
      documented, pseudonomyzed and included in our data bank.&#xD;
&#xD;
      BAL and blood aliquots of 20 control pts without immunosuppression (suffering from lung&#xD;
      diseases) will be collected and tested identically.&#xD;
&#xD;
      Case definitions will be made according to 2008 EORTC/MSG criteria. The duration of the study&#xD;
      will be approximately 24 months&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Unknown status</overall_status>
  <last_known_status>Enrolling by invitation</last_known_status>
  <start_date>August 2012</start_date>
  <completion_date type="Anticipated">December 2020</completion_date>
  <primary_completion_date type="Anticipated">December 2020</primary_completion_date>
  <study_type>Observational</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <observational_model>Cohort</observational_model>
    <time_perspective>Prospective</time_perspective>
  </study_design_info>
  <primary_outcome>
    <measure>Diagnostic certainty</measure>
    <time_frame>6 Weeks</time_frame>
    <description>After 6 weeks the diagnostic performance of the biomarkers and their combination in relation to diagnostic accuracy will be measured</description>
  </primary_outcome>
  <number_of_groups>2</number_of_groups>
  <enrollment type="Anticipated">200</enrollment>
  <condition>Invasive Aspergillosis</condition>
  <condition>Acute Leukemia</condition>
  <condition>Biomarkers</condition>
  <condition>Stem Cell Transplantation</condition>
  <arm_group>
    <arm_group_label>Immunocompromised Patients</arm_group_label>
    <description>Patients with acute leukemia undergoing induction chemotherapy or undergoing allogeneic stem cell transplantation</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Control Group</arm_group_label>
    <description>Patients underdoing bronchoscopy and diagnostic BAL without immunosuppression and without signs of infection (Sarcoidosis, Lung Cancer)</description>
  </arm_group>
  <biospec_retention>Samples With DNA</biospec_retention>
  <biospec_descr>
    <textblock>
      Only fungal DNA is retained and investigated&#xD;
    </textblock>
  </biospec_descr>
  <eligibility>
    <study_pop>
      <textblock>
        Patients suffering from acute leukemia and pts after allogeneic stem cell transplantation&#xD;
        with febrile neutropenia and lung infiltrates diagnosed in a chest CT scan suggestive for&#xD;
        fungal infection will be investigated after pts's informed consent in a multicentre,&#xD;
        prospective trial&#xD;
      </textblock>
    </study_pop>
    <sampling_method>Probability Sample</sampling_method>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  acute leukemia or after allogeneic stem cell transplantation&#xD;
&#xD;
          -  febrile neutropenia&#xD;
&#xD;
          -  lung infiltrates suggestive for fungal infection (halo sign. nodules, air-crescent&#xD;
             sign)&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  missing informed consent&#xD;
&#xD;
          -  other underlying diagnosis&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>18 Years</minimum_age>
    <maximum_age>100 Years</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Dieter Buchheidt, MD</last_name>
    <role>Principal Investigator</role>
    <affiliation>Medical Faculty Mannheim, University of Heidelberg</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>University Hospital Mannheim</name>
      <address>
        <city>Mannheim</city>
        <zip>68167</zip>
        <country>Germany</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>Germany</country>
  </location_countries>
  <reference>
    <citation>Reinwald M, Hummel M, Kovalevskaya E, Spiess B, Heinz WJ, Vehreschild JJ, Schultheis B, Krause SW, Claus B, Suedhoff T, Schwerdtfeger R, Reuter S, Kiehl MG, Hofmann WK, Buchheidt D. Therapy with antifungals decreases the diagnostic performance of PCR for diagnosing invasive aspergillosis in bronchoalveolar lavage samples of patients with haematological malignancies. J Antimicrob Chemother. 2012 Sep;67(9):2260-7. doi: 10.1093/jac/dks208. Epub 2012 Jun 27.</citation>
    <PMID>22740590</PMID>
  </reference>
  <reference>
    <citation>Reinwald M, Spiess B, Heinz WJ, Vehreschild JJ, Lass-Florl C, Kiehl M, Schultheis B, Krause SW, Wolf HH, Bertz H, Maschmeyer G, Hofmann WK, Buchheidt D. Diagnosing pulmonary aspergillosis in patients with hematological malignancies: a multicenter prospective evaluation of an Aspergillus PCR assay and a galactomannan ELISA in bronchoalveolar lavage samples. Eur J Haematol. 2012 Aug;89(2):120-7. doi: 10.1111/j.1600-0609.2012.01806.x. Epub 2012 Jun 22.</citation>
    <PMID>22650156</PMID>
  </reference>
  <reference>
    <citation>Spiess B, Seifarth W, Merker N, Howard SJ, Reinwald M, Dietz A, Hofmann WK, Buchheidt D. Development of novel PCR assays to detect azole resistance-mediating mutations of the Aspergillus fumigatus cyp51A gene in primary clinical samples from neutropenic patients. Antimicrob Agents Chemother. 2012 Jul;56(7):3905-10. doi: 10.1128/AAC.05902-11. Epub 2012 Apr 23.</citation>
    <PMID>22526309</PMID>
  </reference>
  <verification_date>April 2020</verification_date>
  <study_first_submitted>September 26, 2012</study_first_submitted>
  <study_first_submitted_qc>September 26, 2012</study_first_submitted_qc>
  <study_first_posted type="Estimate">September 28, 2012</study_first_posted>
  <last_update_submitted>April 17, 2020</last_update_submitted>
  <last_update_submitted_qc>April 17, 2020</last_update_submitted_qc>
  <last_update_posted type="Actual">April 20, 2020</last_update_posted>
  <responsible_party>
    <responsible_party_type>Principal Investigator</responsible_party_type>
    <investigator_affiliation>Heidelberg University</investigator_affiliation>
    <investigator_full_name>Dieter Buchheidt</investigator_full_name>
    <investigator_title>Professor Dr. Dieter Buchheidt</investigator_title>
  </responsible_party>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Aspergillosis</mesh_term>
  </condition_browse>
  <!-- Results have not yet been posted for this study                                          -->
</clinical_study>


download date: ClinicalTrials.gov processed this data on September 20, 2023
link text: Link to the current ClinicalTrials.gov record.
url: https://clinicaltrials.gov/ct2/show/NCT01695512
org study id: University of Heidelberg
secondary id: Grant Funding number
nct id: NCT01695512
lead sponsor: 
collaborator: 
has dmc: No
The aim of our prospective and multicentre diagnostic study is therefore to elucidate on the
      sensitivity and specificity rates of these serologic markers in combination with molecular
      tools (both an Aspergillus specific and a multifungal PCR based assay), as serologic mark-ers
      are not pathogen-specific, and furthermore to define species-specific cut-off values for BDG
      in BAL samples.

      Additionally, if genomic material of Aspergillus fumigatus is detected by PCR in a clinical
      sample, we investigate fungal DNA for point mutations in the cyp51A gene mediating
      resis-tance against common mould-active triazoles with novel rapid, sensitive and specific,
      non-culture-based PCR-assays and sequencing to optimize antifungal treatment as early as
      pos-sible.
The major problem in managing life threatening invasive fungal infections in patients (pts)
      with acute leukemia and pts after allogeneic hematopoietic stem cell transplantation is the
      lack of sensitive and specific diagnostic tools to identify fungal pathogens reliably and
      early in the course of the disease. Because of deep neutropenia and low platelet count,
      invasive diagnostic procedures are rarely feasible in time, therefore bronchoscopy with BAL
      is the method of choice in diagnosing pulmonary infections, as the sensitivity of
      culture-based methods from blood is low, especially in mould infections. Indirect methods,
      so-called surrogate markers, are becoming increasingly important in this clinical setting.
      These serologic markers, mainly galactomannan (GM) and recently 1(1,3)-β-D-glucan (BDG), have
      been validated in clinical trials for blood samples, however the clinical significance of
      testing BAL samples is up to now only based on retrospective data for GM and has not been
      reported yet for BDG.

      The aim of our prospective and multicentre diagnostic study is therefore to elucidate on the
      sensitivity and specificity rates of these serologic markers in combination with molecular
      tools (both an Aspergillus specific and a multifungal PCR based assay), as serologic markers
      are not pathogen-specific, and furthermore to define species-specific cut-off values for BDG
      in BAL samples.

      Additionally, if genomic material of Aspergillus fumigatus is detected by PCR in a clinical
      sample, we investigate fungal DNA for point mutations in the cyp51A gene mediating resistance
      against common mould-active triazoles with novel rapid, sensitive and specific,
      non-culture-based PCR-assays and sequencing to optimize antifungal treatment as early as
      possible.

      This study aims to improve the early, sensitive and specific diagnosis of invasive pulmonary
      fungal infections and detect azole resistance patterns, it might impact on the prognosis in
      hematologic patients; therefore antifungal treatment data and clinical outcome will be
      recorded.

      Clinical samples (BAL and blood) from approximately 100 pts suffering from acute leukemia and
      pts after allogeneic stem cell transplantation with febrile neutropenia and lung infiltrates
      diagnosed in a chest CT scan suggestive for fungal infection will be investigated after pts's
      informed consent in a multicentre, prospective trial. Pts will undergo standardized
      diagnostic imaging and microbiological procedures for identification of the underlying
      infectious pathogen. BAL and blood samples will be tested additionally for GM, BDG and with a
      diagnostic nested Aspergillus PCR assay, a multifungal DNA-Microarray and an azole resistance
      PCR assay. The molecular assays were established by our group and encompass the detection of
      Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, Candida albicans, Candida
      dubliniensis, Candida glabrata, Candida lusitaniae, Candida tropicalis, Fusarium oxysporum,
      Fusarium solani, Mucor racemosus, Rhizopus microsporus, Scedosporium prolificans,
      Trichosporon asahii and identify the three most common point mutations that confer resistance
      to triazoles like voriconazole or posaconazole.

      Results of other diagnostic means including culture findings as well as patients' clinical
      data (e.g. duration of neutropenia, underlying disease and outcome including follow-up data
      concerning antifungal treatment and mortality attributable to fungal infections) will be
      documented, pseudonomyzed and included in our data bank.

      BAL and blood aliquots of 20 control pts without immunosuppression (suffering from lung
      diseases) will be collected and tested identically.

      Case definitions will be made according to 2008 EORTC/MSG criteria. The duration of the study
      will be approximately 24 months
observational model: Cohort
time perspective: Prospective
measure: Diagnostic certainty
time frame: 6 Weeks
description: After 6 weeks the diagnostic performance of the biomarkers and their combination in relation to diagnostic accuracy will be measured
arm group label: Immunocompromised Patients
description: Patients with acute leukemia undergoing induction chemotherapy or undergoing allogeneic stem cell transplantation
arm group label: Control Group
description: Patients underdoing bronchoscopy and diagnostic BAL without immunosuppression and without signs of infection (Sarcoidosis, Lung Cancer)
Only fungal DNA is retained and investigated
study pop: 
sampling method: Probability Sample
criteria: 
gender: All
minimum age: 18 Years
maximum age: 100 Years
healthy volunteers: Accepts Healthy Volunteers
last name: Dieter Buchheidt, MD
role: Principal Investigator
affiliation: Medical Faculty Mannheim, University of Heidelberg
facility: 
country: Germany
citation: Reinwald M, Hummel M, Kovalevskaya E, Spiess B, Heinz WJ, Vehreschild JJ, Schultheis B, Krause SW, Claus B, Suedhoff T, Schwerdtfeger R, Reuter S, Kiehl MG, Hofmann WK, Buchheidt D. Therapy with antifungals decreases the diagnostic performance of PCR for diagnosing invasive aspergillosis in bronchoalveolar lavage samples of patients with haematological malignancies. J Antimicrob Chemother. 2012 Sep;67(9):2260-7. doi: 10.1093/jac/dks208. Epub 2012 Jun 27.
PMID: 22740590
citation: Reinwald M, Spiess B, Heinz WJ, Vehreschild JJ, Lass-Florl C, Kiehl M, Schultheis B, Krause SW, Wolf HH, Bertz H, Maschmeyer G, Hofmann WK, Buchheidt D. Diagnosing pulmonary aspergillosis in patients with hematological malignancies: a multicenter prospective evaluation of an Aspergillus PCR assay and a galactomannan ELISA in bronchoalveolar lavage samples. Eur J Haematol. 2012 Aug;89(2):120-7. doi: 10.1111/j.1600-0609.2012.01806.x. Epub 2012 Jun 22.
PMID: 22650156
citation: Spiess B, Seifarth W, Merker N, Howard SJ, Reinwald M, Dietz A, Hofmann WK, Buchheidt D. Development of novel PCR assays to detect azole resistance-mediating mutations of the Aspergillus fumigatus cyp51A gene in primary clinical samples from neutropenic patients. Antimicrob Agents Chemother. 2012 Jul;56(7):3905-10. doi: 10.1128/AAC.05902-11. Epub 2012 Apr 23.
PMID: 22526309
responsible party type: Principal Investigator
investigator affiliation: Heidelberg University
investigator full name: Dieter Buchheidt
investigator title: Professor Dr. Dieter Buchheidt
mesh term: Aspergillosis