Diagnosis and treatment of invasive aspergillosis

Abstract

Methods for diagnosing, treating, and monitoring the treatment of invasive aspergillosis (IA) are described. The methods can include detecting the presence of one or more volatile organic compounds (VOCs) in the breath of subjects suspected of having IA.

What is claimed is: 
     
         1 . A method for diagnosing a subject with invasive aspergillosis (IA), the method comprising:
 obtaining a sample comprising breath of a subject or suspected of comprising  Aspergillus  isolated from a subject;   detecting the presence in the sample of one, two, three, or more volatile organic compounds (VOCs) produced by the  Aspergillus  species in a sample comprising breath from the subject or headspace from a culture suspected of comprising  Aspergillus  isolated from the subject, wherein the VOCs are selected from the group consisting of camphene, α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, and trans-geranylacetone; and   diagnosing a subject as having IA when there are one, two, three or more of the VOCs present in the sample.   
     
     
         2 . The method of  claim 1 , comprising:
 (i) detecting the presence in the sample of α-trans-bergamotene, β-trans-bergamotene, and trans-geranylacetone; and   diagnosing a subject who has all three of α-trans-bergamotene, β-trans-bergamotene, and trans-geranylacetone in the sample as having IA, e.g., wherein the sample is a sample of a subject's breath; or   (ii) detecting the presence in the sample of camphene, α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, and trans-geranylacetone; and   diagnosing a subject who has all of camphene, α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, and trans-geranylacetone, e.g., wherein the sample comprises headspace from a culture.   
     
     
         3 . A method of treating a subject who has invasive aspergillosis (IA), the method comprising:
 obtaining a sample comprising breath of a subject or headspace from a culture suspected of comprising  Aspergillus  isolated from a subject;   detecting the presence in the sample of one, two, three, or more VOCs selected from the group consisting of camphene, α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, and trans-geranylacetone; and   administering an antifungal treatment to a subject who has one, two, three, or more of camphene, α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, and trans-geranylacetone, e.g., α-trans-bergamotene, β-trans-bergamotene, and trans-geranylacetone in their breath or camphene, α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, and trans-geranylacetone in headspace from a culture.   
     
     
         4 . The method of  claim 3 , wherein the treatment comprises administration of one or more doses of one or more antifungal compounds. 
     
     
         5 . A method of detecting the presence of an  Aspergillus fumigatus, A. terreus , or  A. calidoustus  infection in a subject, the method comprising:
 obtaining a sample comprising breath of a subject, or headspace from a culture suspected of comprising  Aspergillus  isolated from a subject;   determining the presence of one, two, three, or more, e.g., all, of the VOCs selected from the group consisting of camphene, α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, trans-geranylacetone, elixene, alpha-santalene, beta-elemene, acoradien, chamigrene, 1,5,9-trimethyl cyclododecatriene, 9-decene-2-one and beta-sesquiphellandrene in the sample, wherein:
 the presence of one, two, three, or more, e.g., all, of camphene, α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, and trans-geranylacetone indicates the presence of an  A. fumigatus  infection; 
 the presence of one, two, three, or more, e.g., all, of elixene, alpha-santalene, beta-elemene, acoradien, chamigrene, and 1,5,9-trimethyl cyclododecatriene indicates the presence of an  A. terreus  infection; and 
 the presence of one or both of 9-decene-2-one and beta-sesquiphellandrene indicates the presence of an  A. calidoustus  infection in the subject. 
   
     
     
         6 . The method of  claim 5 , further comprising selecting, and optionally administering, a therapy comprising an azole, e.g., voriconazole, for a subject who has an  A. fumigatus  or  A. terreus  infection; or
 selecting, and optionally administering, a therapy comprising amphotericin B (AMB), e.g., D-AMB or a lipid formulation of AMB, for a subject who has an  A. calidoustus  infection.   
     
     
         7 . A method of monitoring efficacy of a treatment for invasive aspergillosis (IA) in a subject, the method comprising:
 determining a first level of one, two, three, or more volatile organic compounds (VOCs) produced by the  Aspergillus  species in a sample comprising breath from the subject or headspace from a culture suspected of comprising  Aspergillus  isolated from the subject, wherein the VOCs are selected from the group consisting of camphene, α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, trans-geranylacetone, elixene, alpha-santalene, beta-elemene, acoradien, chamigrene, 1,5,9-trimethyl cyclododecatriene, 9-decene-2-one and beta-sesquiphellandrene, in the subject;   administering a treatment for IA to the subject;   determining a second level of the VOCs in a sample obtained after administration of the treatment to the subject; and   comparing the first and second levels of VOCs, wherein a decrease in the VOCs indicates that the treatment has been effective in treating the IA in the subject, and an increase or no change indicates that the treatment has not been effective in treating the IA in the subject.   
     
     
         8 . The method of  claim 6 , wherein the treatment comprises administration of one or more doses of one or more antifungal compounds. 
     
     
         9 . A method of identifying a candidate compound for the treatment of IA, the method comprising:
 providing a test culture comprising one or more  Aspergillus  species;   detecting a baseline level of fungal VOCs in the headspace of the culture in the absence of the test compound, wherein the VOCs are selected from the group consisting of camphene, α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, trans-geranylacetone, elixene, alpha-santalene, beta-elemene, acoradien, chamigrene, 1,5,9-trimethyl cyclododecatriene, 9-decene-2-one and beta-sesquiphellandrene, in the subject;   contacting the test culture with a test compound;   determining a second level of the VOCs in a the test culture;   comparing the second level of VOCs to the baseline level; and   identifying a test compound that decreases levels of fungal VOCs in the test culture as a candidate compound for the treatment of IA.   
     
     
         10 . A method of detecting the presence of an  Aspergillus fumigatus, A. terreus , or  A. calidoustus  infection in a culture, the method comprising:
 obtaining a sample from the culture, e.g., gas from the headspace of the culture;   determining the presence of one, two, three, or more, e.g., all, of the VOCs selected from the group consisting of camphene, α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, trans-geranylacetone, elixene, alpha-santalene, beta-elemene, acoradien, chamigrene, 1,5,9-trimethyl cyclododecatriene, 9-decene-2-one and beta-sesquiphellandrene in the sample, wherein:
 the presence of one, two, three, or more, e.g., all, of α-pinene, β-pinene, limonene, α-trans-bergamotene, β-trans-bergamotene, trans-geranylacetone indicates the presence of  A. fumigatus  in the culture; 
 the presence of one, two, three, or more, e.g., all, of elixene, alpha-santalene, beta-elemene, acoradien, chamigrene, and 1,5,9-trimethyl cyclododecatriene indicates the presence of  A. terreus  in the culture; and 
 the presence of one or both of 9-decene-2-one and beta-sesquiphellandrene indicates the presence of  A. calidoustus  infection in the culture. 
   
     
     
         11 . The method of any of  claims 1 - 10 , wherein determining the presence of a VOC comprises assaying the sample to detect the presence the VOC. 
     
     
         12 . The method of  claim 11 , wherein assaying the sample to detect the presence the VOC comprises using a gas chromatography (GC) or spectrometry method. 
     
     
         13 . The method of  claim 12 , wherein the spectrophotometry method is mobility spectrometry (IMS) or differential mobility spectrometry (DMS). 
     
     
         14 . The method of any of  claims 1 - 10 ,  12 , or  13 , wherein the subject is a human.\ 
     
     
         15 . The method of  claim 11 , wherein the subject is a human. 
     
     
         16 . The method of  claims 4  or  8 , wherein the antifungal compound is an amphotericin B formulation; an azole antifungal compound; or an echinocandin antifungal compound.