![[logo] US EPA](https://webarchive.library.unt.edu/eot2008/20081109144227im_/http://www.epa.gov/epafiles/images/logo_epaseal.gif){kind=logo}
EPA Technology for Mold Identification and Enumeration
The presence of molds in homes, schools and business is of growing concern in the United States. To begin to understand the significance of exposure to these organisms, a standardized technology is needed for their identification and quantification. In May of 2002, the EPA was granted patent 6,387,652 which describes mold DNA sequences that makes their identification and quantification possible. This relatively inexpensive technology has been licensed by firms in the U.S., Germany and the United Kingdom (below).The patented technology teaches the best available DNA sequences to use for over 130 of the major indoor air fungi. It is particularly useful, in that a a licensed company can employ a technician or robotic system to analyze samples and obtain highly reproducible results in only a few hours -- eliminating the need for plating and culturing or identifying and counting. Other analytical techniques may take days or week to complete. Most typical sampling systems can be adapted for use with this technology. The EPA, through cooperative research and development agreements (CRADAs), instructs company staff on its utilization. This training is completed in two or three days.
Literature references pertinent to this technology can be found at the end of this page.
Companies Licensed to Utilize Mold Technology (as of 6/20/07)
| AEMTEK, Inc. Contact: Steven Huang 46309 Warm Springs Blvd, Suite A Fremont, CA 94539 Telephone: 510-979-1979 ext. 302 |
Genetic ID Contact: Bernd Schoel 501 Domick Dr. Fairfield, IA Telephone: 1-641-472-9979 ext. 124 |
| Aerotech Laboratories, Inc. Contact: Janice Jones 1501 W. Knudsen Dr. Phoenix, AZ 85027 Telephone: 800-651-4802 ext.1027 |
Healthy Office Co. UK Limited Contact: Richard Stott 28 John St., Hull East Yorkshire HU2 8DH, United Kingdom Telephone: +44 (0) 1482 328410 |
| Ambient Group, Inc. Contact:Sean Wallace 470 7th Ave. 12th Floor New York, NY 10018 Telephone: 212-944-4615 |
Microbac Laboratories Contact: Robert Brooks 505 East Broadway Maryville, TN 37804 Telephone: 865-977-1200 |
| CENAS AG Contact: Alexandra Kaetzke E.-C-Baumann-Str 20 95326 Kulmbach Deutschland Telephone: 0 92 21 / 8 27 61 30 |
Microbial Insights, Inc Contact: Greg Davis 2340 Stock Creek Blvd. Rockford, TN 37853-3044 Telephone: 865-573-8188 |
| Clean Air Labs Contact: James Campbell 228 Midway Ln, Suite B Oak Ridge, TN 37830 Telephone: 865-483-9793 |
Molecular World, Inc. Contact: Curtis Hildebrandt Cumberland Park 1-South Cumberland St. Thunder Bay, ON P7B 2T1 Telephone: 1-807-344-7666 Toll Free: 1-877-665-9753 |
| EMSL Analytical, Inc. Contact: Quanyi "Charlie" Li 107 Haddon Ave. Westmont, NJ 08108 Telephone: 800-220-3675 ext. 1283 |
Mycometrics, LLC Contact: King-Teh Lin 11 Deer Park Dr., Suite 210 Monmouth Junction, NJ 08852 Telephone: 732-355-9018 |
|
Forensic Analytical |
Pure Earth Environmental Laboratory, Inc. Contact: Theodore J. Passon, Jr. 7184 N. Park Drive Pennsauken, NJ 08110 Telephone: 856-486-1177 ext. 222 |
|
Galson Laboratories |
P & K Microbiology Services Contact: Michael Berg 1936 Olney Ave. Cherry Hill, NJ 08003 Telephone: 856-489-4455 |
Primers and Probes for Target Species
Absidia | Acremonium | Alternaria | Aspergillus | Aureobasidium | Candida | Chaetomium | Cladosporium | Emericella | Eurotium | Epicoccum | Fusarium | Geotrichum | Memnoniella | Mucor | Myrothecium | Paecilomyces | Penicillium | Rhizomucor | Rhizopus | Scopulariopsis | Stachybotrys | Trichoderma | Ulocladium | Wallemia | Universial Penicillium, Aspergillus, Paecilomyces |
Absidia corymbifera (assay name: Acory)
Forward Primer NS92F: 5'-CACCGCCCGTCGCTAC
Reverse Primer AcoryR1: 5'-GCAAAGCGTTCCGAAGGACA
Probe AcoryP1: 5'-ATGGCACGAGCAAGCATTAGGGACG
Acremonium strictum (assay name: Astrc)
Forward Primer AstrcF1: 5'-CAACCCATTGTGAACTTACCAAAC
Reverse Primer AstrcR1: 5'-CGCCCCTCAGAGAAATACGATT
Probe AstrcP1: 5'-TCAGCGCGCGGTGGCCTC
Alternaria alternata (assay name: Aaltr)
Forward Primer AaltrF1: 5'-GGCGGGCTGGAACCTC
Reverse Primer AltrR1-1: 5'-GCAATTACAAAAGGTTTATGTTTGTCGTA
Probe AaltrP1: 5'-TTACAGCCTTGCTGAATTATTCACCCTTGTCTTT
Aspergillus auricomus (assay name: Aauri)
Forward Primer AcircF1: 5'-ATTACTGAGTGAGGGTCCCTCG
Reverse Primer AauriR1: 5'-GGCGGCCGCGTAAAC
Probe AcircP1: 5'-CCCGCCGAAGCAACAAGGTACG
Aspergillus caespitosus (assay name: Acaes)
Forward Primer AcaesF1: 5'-CTCCCACCCGTGAATACCTT
Reverse Primer AcaesR1: 5'-GGCTCAGACGCAACTCTACAAT
Probe AcaesP1: 5'-CACTGTTGCTTCGGCGAGGAGCC
Aspergillus candidus (assay name: Acand3)
Forward Primer AcandF1: 5'-TTACCGAGTGAGGGTTTCTCTGA
Reverse Primer AcandR2: 5'-CCCGCCGAAGCAACAG
Probe* PenAspP1mgb: 5'-CCAACCTCCCACCCGTG
Aspergillus carbonarius (assay name: Acarb)
Forward Primer AcarbF1-1: 5'-CTTTGGGCCCAACCTCCtAC
Reverse Primer AcarbR1: 5'-CCCGAGGGGCAGAGATG
Probe AspP1: 5'-TTGCTTCGGCGGGCCC
Aspergillus cervinus (assay name: Acerv)
Forward Primer AcervF1: 5'-CCACCCGTGCTATTGTACCTTT
Reverse Primer AcervR1-1: 5'-CAACTCAGACTGCAATTCAGAACtGT
Probe AspP1: 5'-TTGCTTCGGCGGGCCC
Aspergillus clavatus/giganteus (assay name: Aclav)
Forward Primer AclavF1: 5'-CCCGCCGTCTTCGGA
Reverse Primer AclavR1: 5'-CCGTTGTTGAAAGTTTTAACTGATTATG
Probe AfumiP1: 5'-CCCGCCGAAGACCCCAACATG
Aspergillus flavus/oryzae (assay name: Aflav)
Forward Primer AflavF1: 5'-CGAGTGTAGGGTTCCTAGCGA
Reverse Primer AflavR1: 5'-CCGGCGGCCATGAAT
Probe AflavP1: 5'-TCCCACCCGTGTTTACTGTACCTTAGTTGCT
Aspergillus fumigatus, Neosartorya fischeri (assay name: Afumi)
Forward Primer AfumiF1: 5'-GCCCGCCGTTTCGAC
Reverse Primer AfumiR1: 5'-CCGTTGTTGAAAGTTTTAACTGATTAC
Probe AfumiP1: 5'-CCCGCCGAAGACCCCAACATG
Aspergillus flavipes (assay name: Aflvp2)
Forward Primer AflvpF1: 5'-CCACCCGTGACTACTGTACCAC
Reverse Primer AflvpR2: 5'-AGGCTTTCAGAAACAGTGTTCG
Probe AspP1: 5'-TTGCTTCGGCGGGCCC
Aspergillus niger/awamori/foetidus/phoenicis (assay name: Anigr)
Forward Primer AnigrF1: 5'-GCCGGAGACCCCAACAC-3'
Reverse Primer AnigrR1: 5'-TGTTGAAAGTTTTAACTGATTGCATT-3'
Probe AnigrP1: 5'-AATCAACTCAGACTGCACGCTTTCAGACAG
Aspergillus niveus (assay name: Anive)
Forward Primer AniveF1: 5'-ACCCGTGCCTATTGTACCCT
Reverse Primer AniveR1: 5'-TGCAAACAATCACACTCAGACAC
Probe AspP1: 5'-TTGCTTCGGCGGGCCC
Aspergillus ochraceus/ostianus (assay name: Aochr1)
Forward Primer AochrF1: 5'-AACCTCCCACCCGTGTATACC-3'
Reverse Primer AochrR1: 5'-CCGGCGAGCGCTGTG-3'
Probe AochrP1: 5'-ACCTTGTTGCTTCGGCGAGCCC
Aspergillus paradoxus (assay name: Apard)
Forward Primer ApardF1: 5'-CGGGGGGCTTACGCT
Reverse Primer ApardR1-1: 5'-GACTGCAACTTCATACAGAGTTGGT
Probe PenP2: 5'-CGCGCCCGCCGAAGACA
Aspergillus parasiticus/sojae (assay name: Apara)
Forward Primer AflavF1: 5'-CGAGTGTAGGGTTCCTAGCGA-3'
Reverse Primer AparaR3: 5'-GCCCGGGGCTGACG
Probe AflavP1: 5'-TCCCACCCGTGTTTACTGTACCTTAGTTGCT
Aspergillus penicillioides (assay name: Apeni2)
Forward Primer ApeniF2: 5'-CGCCGGAGACCTCAACC
Reverse Primer ApeniR2: 5'-TCCGTTGTTGAAAGTTTTAACGA
Probe ApeniP2: 5'-TGAACACTGTCTGAAGGTTGCAGTCTGAGTATG
Aspergillus puniceus (assay name: Apuni)
Forward Primer ApuniF1: 5'-CATTACCGAGTGCAGGCCTC
Reverse Primer AustsR1: 5'-GCCGAAGCAACGTTGGTC
Probe ApuniP1: 5'-CCCCACAGGCAGGCCTAACCTC
Aspergillus restrictus/caesillus/conicus (assay name: Arest)
Forward Primer ArestF1: 5'-GGGCCCGCCTTCAT-3'
Reverse Primer ArestR1: 5'-GTTGTTGAAAGTTTTAACGATTTTTCT
Probe ArestP1: 5'-CCCGCCGGAGACTCCAACATTG
Aspergillus sclerotiorum (assay name: Asclr)
Forward Primer AcircF1: 5'-ATTACTGAGTGAGGGTCCCTCG
Reverse Primer AsclrR1: 5'-CCTAGGGAGGGGGGTTTGA
Probe AcircP1: 5'-CCCGCCGAAGCAACAAGGTACG
Aspergillus sydowii (assay name: Asydo3)
Forward Primer AsydoF1-1: 5'-CAACCTCCCACCCGAGAA
Reverse Primer AversR1-1: 5'-CCATTGTTGAAAGTTTTGACTGATcTTA
Probe versP1: 5'-AGACTGCATCACTCTCAGGCATGAAGTTCAG
Aspergillus tamarii (assay name: Atama2)
Forward Primer AflavF1: 5'-CGAGTGTAGGGTTCCTAGCGA
Reverse Primer AtamaR2: 5'-CCCGGCGGCCTTAAAG
Probe AflavP1: 5'-TCCCACCCGTGTTTACTGTACCTTAGTTGCT
Aspergillus terreus (assay name: Aterr2)
Forward Primer AterrF1-1: 5'-ATCATTACCGAGTGCGTGTCTTTA
Reverse Primer AterrR2: 5'-CCCGCCGAAGCAACAAG
Probe AterrP2: 5'-CCCAACCTCCCACCCGTGACTATTG
Aspergillus unguis (assay name: Aungu)
Forward Primer AunguF1: 5'-CAACCTCCCACCCTTGAATACT
Reverse Primer AunguR1: 5'-TCACTCTCAGGCATGAAGTTCAG
Probe AcaesP1: 5'-CACTGTTGCTTCGGCGAGGAGCC
Aspergillus ustus (assay name: Austs2)
Forward Primer AustsF2-1: 5'-AAGGATCATTACCGAGTGCAtGT
Reverse Primer AustsR1: 5'-GCCGAAGCAACGTTGGTC
Probe AustsP1: 5'-CCCCCGGGCAGGCCTAACC
Aspergillus versicolor (assay name: Avers2-2)
Forward Primer AversF2 (5x): 5'-CGGCGGGGAGCCCT
Reverse Primer AversR1-1: 5'-CCATTGTTGAAAGTTTTGACTGATcTTA
Probe versP1: 5'-AGACTGCATCACTCTCAGGCATGAAGTTCAG
Aspergillus wentii (assay name: Awent)
Forward Primer AwentF1: 5'-CATTACCGAGTGAGGACCTAACC
Reverse Primer AwentR1: 5'-CGGCGGCCACGAAT
Probe AcircP1: 5'-CCCGCCGAAGCAACAAGGTACG
Aureobasidium pullulans (assay name:
Apull)
Forward Primer ApullF1: 5'-GATCATTAAAGAGTAAGGGTGCTCA
Reverse Primer ApullR1: 5'-GCTCGCCTGGGACGAATC
Probe ApullP1: 5'-CGCCCGACCTCCAACCCTTTG
Candida albicans (assay name: Calbi)
Forward Primer CalbF1: 5'-CTTGGTATTTTGCATGTTGCTCTC
Reverse Primer CalbR1: 5'-GTCAGAGGCTATAACACACAGCAG
Probe CalbP1: 5'-TTTACCGGGCCAGCATCGGTTT
Candida dubliniensis (assay name: Cdubl)
Forward Primer CdubF1: 5'-AGATCAGACTTGGTATTTTGCAAGTTA
Reverse Primer CdubR1: 5'-TAGGCTGGCAGTATCGTCAGA
Probe CdubP1: 5'-TTTACCGGGCCAGCATCGGTTT
Candida glabrata (assay name: Cglab)
Forward Primer Cglab F1: 5'-GCGCCCCTTGCCTCTC
Reverse Primer Cglab R1: 5'-CCCAGGGCTATAACACTCTACACC
Probe Cglab P1: 5'-TGGGCTTGGGACTCTCGCAGC
Candida (assay name: Pichia) guilliermondii (assay name:
Cguil)
Forward Primer CguiF1: 5'-CCTTCGTGGCGGGGTG
Reverse Primer CguiR1: 5'-GCAGGCAGCATCAACGC
Probe CguiP1: 5'-CCGCAGCTTATCGGGCCAGC
Candida haemulonii, svar. 1 (assay name: Chaem1)
Forward Primer ChaeF1: 5'-GGAGCGACAACGAGCAGTC
Reverse Primer ChaeR1: 5'-AGGAGCCAGAAAGCAAGACG
Probe ChaeP1: 5'-ATGTAGTACAGCCCTCTGGGCTGTGCA
Candida haemulonii, svar. 2 (assay name: Chaem2)
Forward Primer Cha2F1: 5'-ATCGGGTGGAGCGGAACT
Reverse Primer Cha2R1: 5'-CGAAGCAGGAACCATCTGAGA
Probe Cha2P1: 5'-AAGTGGGAGCTGATGTAGCAACCCCC
Candida krusei (assay name: Ckrus)
Forward Primer CkruF1: 5'-CTCAGATTTGAAATCGTGCTTTG
Reverse Primer CkruR1: 5'-GGGGCTCTCACCCTCCTG
Probe CkruP1: 5'-CACGAGTTGTAGATTGCAGGTTGGAGTCTG
Candida lipolytica (assay name: Clipo)
Forward Primer ClipF1: 5'-TAGCGAGACGAGGGTTACAAATG
Reverse Primer ClipR1: 5'-CGTCGGTGGCAGTGTGGA
Probe ClipP1: 5'-CCTTCGGGCGTTCTCCCCTAACC
Candida lusitaniae (assay name: Clusi)
Forward Primer Clus F2: 5'-GGGAATTGTAATTTGAAGGTTTCGT-3'
Reverse Primer Clus R2: 5'-GTCGGCGTGCGCCATA-3'
Probe Clus P2: 5'-TCTGAGTCGGCCGCGCCC-3'
Candida maltosa (assay name: Cmalt)
Forward Primer CmalF1: 5'-GGCCAGCATCAGTTTGGAC
Reverse Primer CmalR1: 5'-TCTAGACTGGCAGTATCGACAGTG
Probe CmalP1: 5'-TAGGACAATTGCGGTGGAATGTGGC
Candida parapsilosis (assay name: Cpara)
Forward Primer CparF1: 5'-GATCAGACTTGGTATTTTGTATGTTACTCTC
Reverse Primer CparR1: 5'-CAGAGCCACATTTCTTTGCAC
Probe CparP1: 5'-CCTCTACAGTTTACCGGGCCAGCATCA
Candida sojae (assay name: Csoja)
Forward Primer CsojF1: 5'-CGGTTGTGTGTTATAGCCTTCGTA
Reverse Primer CsojR1: 5'-ATCATTATGCCAACATCCTAGGTAAT
Probe CtropP2: 5'-CGCAGTCCTCAGTCTAGGCTGGCAG
Candida tropicalis (assay name: Ctrop)
Forward Primer CtropF1: 5'-GCGGTAGGAGAATTGCGTT
Reverse Primer CtropR2: 5'-TCATTATGCCAACATCCTAGGTTTA
Probe CtropP2: 5'-CGCAGTCCTCAGTCTAGGCTGGCAG
Candida viswanathii (assay name: Cvisw)
Forward Primer CvisF1: 5'-CGGCAGGACAATCGCGT
Reverse Primer CvisR1: 5'-TCTAGGCTGGCAGTATCCACG
Probe CvisP1: 5'-AATGTGGCACGGCCTCGGC
Candida zeylanoides (assay name: Czeyl)
Forward Primer Czey F1: 5'-GTTGTAATTTGAAGAAGGTAACTTTGATT
Reverse Primer Czey R1: 5'-GACTCTTCGAAAGCACTTTACATGG
Probe Czey P1: 5'-CCTTGGAACAGGACGTCACAGAGGGT
Chaetomium globosum (assay name: Cglob)
Forward Primer CglobF1: 5'-CCGCAGGCCCTGAAAAG
Reverse Primer CglobR1: 5'-CGCGGCGCGACCA
Probe CglobP1: 5'-AGATGTATGCTACTACGCTCGGTGCGACAG
Cladosporium cladosporioides, svar.
1 (assay name: Cclad1)
Forward Primer Cclad1F1: 5'-CATTACAAGTGACCCCGGTCTAAC
Reverse Primer CcladR1: 5'-CCCCGGAGGCAACAGAG
Probe CcladP1: 5'-CCGGGATGTTCATAACCCTTTGTTGTCC
Cladosporium cladosporioides, svar. 2 (assay name: Cclad2)
Forward Primer Cclad2F1: 5'-TACAAGTGACCCCGGCTACG
Reverse Primer CcladR1: 5'-CCCCGGAGGCAACAGAG
Probe CcladP1: 5'-CCGGGATGTTCATAACCCTTTGTTGTCC
Cladosporium herbarum (assay name: Cherb)
Forward Primer CherbF1: 5'-AAGAACGCCCGGGCTT
Reverse Primer CherbR1: 5'-CGCAAGAGTTTGAAGTGTCCAC
Probe CherbP1: 5'-CTGGTTATTCATAACCCTTTGTTGTCCGACTCTG
Cladosporium sphaerospermum (assay name: Cspha)
Forward Primer CsphaF1: 5'-ACCGGCTGGGTCTTTCG
Reverse Primer CsphaR1: 5'-GGGGTTGTTTTACGGCGTG
Probe CsphaP1: 5'-CCCGCGGCACCCTTTAGCGA
Emericella (Aspergillus) nidulans/rugulosa/quadrilineata
(assay name: Anidu2)
Forward Primer AversF1: 5'-CAACCTCCCACCCGTGAC
Reverse Primer AniduR1-1: 5'-CCATTGTTGAAAGTTTTGACTGATaTGT
Probe versP1: 5'-AGACTGCATCACTCTCAGGCATGAAGTTCAG
Emericella (Aspergillus) variecolor (assay name: Avari)
Forward Primer AversF2: 5'-CGGCGGGGAGCCCT
Reverse Primer AvariR1-1: 5'-CCATTGTTGAAAGTTTTGACTAAATTCTC
Probe AvariP1: 5'-CATCACTACAGGCATGAAGTTCGGTGGT
Eurotium (Aspergillus) amstelodami/chevalieri/herbariorum/rubrum/repens
(assay name: Eamst)
Forward Primer EamstF1: 5'-GTGGCGGCACCATGTCT
Reverse Primer EamstR1: 5'-CTGGTTAAAAAGATTGGTTGCGA
Probe EamstP1: 5'-CAGCTGGACCTACGGGAGCGGG
Epicoccum nigrum (assay name: Enigr)
Forward Primer EnigrF1: 5'-TTGTAGACTTCGGTCTGCTACCTCTT
Reverse Primer EnigrR1: 5'-TGCAACTGCAAAGGGTTTGAAT
Probe EnigrP1: 5'-CATGTCTTTTGAGTACCTTCGTTTCCTCGGC
Fusarium solani f.sp. batatas MP
II (Fsola2)
Forward Primer FsolaF2: 5'-CGCCGCAGCTTCCAAT
Reverse Primer FsolaR2: 5'-GATTCGAGGTCAACGTTCTAAAAGT
Probe* FsolaP1mgb: 5'-CGTAGTAGCTAACACCTC
Fusarium solani f.sp. cucurbitae MP I (Fsola1)
Forward Primer FsolaF1: 5'-CAGCCGCAGCTTCCAGT
Reverse Primer FsolaR1-1: 5'-CGTGGCCGAGCCtCTG
Probe* FsolaP1mgb: 5'-CGTAGTAGCTAACACCTC
Fusarium solani f.sp. mori MP III, Fusarium solani
f.sp. cucurbitae MP V, Fusarium solani f.sp. pisi
MP VI, Fusarium solani f.sp. robiniae MP VII (Fsola3)
Forward Primer FsolaF3: 5'-CGCCGCAGCTTCCATC
Reverse Primer FsolaR3: 5'-GATTCGAGGTCAACTTCAGAAGAG
Probe* FsolaP1mgb: 5'-CGTAGTAGCTAACACCTC
Fusarium solani f.sp. xanthoxyli MP IV (Fsola4)
Forward Primer FsolaF4: 5'-CGCCGCAGCTTCCATT
Reverse Primer FsolaR4-1: 5'-GATTCGAGGTCAACTTTCAGAtGT
Probe* FsolaP1mgb: 5'-CGTAGTAGCTAACACCTC
Geotrichum candidum strain UAMH
7863 (assay name: Geo - reference strain only)
Forward Primer GeoF1: 5'-GATATTTCTTGTGAATTGCAGAAGTGA
Reverse Primer GeoR1: 5'-TTGATTCGAAATTTTAGAAGAGCAAA
Probe GeoP1: 5'-CAATTCCAAGAGAGAAACAACGCTCAAACAAG
Geotrichum candidum (assay name: Gcand2)
Forward Primer NS92F: 5'-CACCGCCCGTCGCTAC
Reverse Primer GcandR1: 5'-AGAAAAGTTGCCCTCTCCAGTT
Probe GeoP2: 5'-TCAATCCGGAAGCCTCACTAAGCCATT
Geotrichum klebahnii (assay name: Gkleb1)
Forward Primer GklebF1: 5'-GGGCGACTTTTCCGGC
Reverse Primer GklebR2: 5'-TGGCACAAATTCTCCTCTAATTTATTTA
Probe GklebP1: 5'-AAGCTAGTCAAACTTGGTCATTTAGAGGAAGTAAAAGTC
Memnoniella echinata (assay name: Mem)
Forward Primer StacF4 5'-TCCCAAACCCTTATGTGAACC
Reverse Primer MemR1: 5'-TGTTTATACCACTCAGACGATACTCAAGT
Probe MemP1: 5'-CTCGGGCCCGGAGTCAGGC
Mucor amphibiorum/circinelloides/hiemalis/indicus/mucedo/racemosus/ramosissimus
and Rhizopus azygosporus/homothalicus/microsporus/oligosporus/oryzae
(assay name: Muc1)
Forward Primer NS92F: 5'-CACCGCCCGTCGCTAC
Reverse Primer MucR1-1: 5'-CCTAGTTTGCCATAGTTCTCAGCAG
Probe MucP1: 5'-CCGATTGAATGGTTATAGTGAGCATATGGGATC
Myrothecium verrucaria (assay name:
Mverr)
Forward Primer MyroF1: 5'-AGTTTACAAACTCCCAAACCCTTT
Reverse Primer MyroR1: 5'-GTGTCACTCAGAGGAGAAAACCA
Probe MyroP1: 5'-CGCCTGGTTCCGGGCCC
Paecilomyces lilacinus (assay name:
Plila)
Forward Primer PlilaF1: 5'-CCCACTGTGAACCTTACCTCAG
Reverse Primer PlilaR1: 5'-GCTTGTGCAACTCAGAGAAGAAAT
Probe PlilaP1: 5'-CCGCCCGCTGGGCGTAATG
Paecilomyces variotii (assay name: Pvari2)
Forward Primer PvariF2: 5'-CGAAGACCCCTGGAACG
Reverse Primer PvariR1: 5'-GTTGTTGAAAGTTTTAATTGATTGATTGT
Probe PvariP1: 5'-CTCAGACGGCAACCTTCCAGGCA
Penicillium aethiopicum (assay name:
Paeth)
Forward Primer PaethF1-1: 5'-CGGGGGGCTCtCGCT
Reverse Primer PchryR1-1: 5'-GAAAGTTTTAAATAATTTATATTTTCACTCAGAgTA
Probe PenP2: 5'-CGCGCCCGCCGAAGACA
Penicillium atramentosum (assay name: Patra)
Forward Primer PgrisF1-1: 5'-ACCTGCGGAAGGATCATTtCT
Reverse Primer PatraR1: 5'-CCCCGGCGGCCATA
Probe PenP3: 5'-TCCAACCTCCCACCCGTGTTTATTT
Penicillium aurantiogriseum/freii/polonicum/tricolor/viridicatum
(assay name: PenGrp1A)
Forward Primer PexpaF1-1: 5'-TTACCGAGTGAGGGCCgTT
Reverse Primer PauraR6: 5'-CCCGGCGGCCAGTA
Probe PenP3: 5'-TCCAACCTCCCACCCGTGTTTATTT
Penicillium aurantiogriseum/freii/hirsutum/polonicum/tricolor/viridicatum/verrucosum,
svar. 2 (assay name: PenGrp1)
Forward Primer PauraF1: 5'-CGGGCCCGCCTTTAC
Reverse Primer PauraR1-1: 5'-GAAAGTTTTAAATAATTTATATTTTCACTCAGAGTT
Probe PenP2: 5'-CGCGCCCGCCGAAGACA
Penicillium brevicompactum/stoloniferum (assay name: Pbrev)
Forward Primer PbrevF3: 5'-GGCGAGCCTGCCTTTTG
Reverse Primer PenR2: 5'-GATCCGTTGTTGAAAGTTTTAAATAATTTATA
Probe PbrevP2: 5'-CTCGCCGAAGACACCTTAGAACTCTGTCTGA
Penicillium canescens (assay name: Pcane2)
Forward Primer PcaneF1: 5'-TTACCGAGCGAGAATTCTCTGA
Reverse Primer PcaneR2-1: 5'-CCGCCGAAGCAACAAGtTAC
Probe* PenAspP1mgb: 5'-CCAACCTCCCACCCGTG
Penicillium chrysogenum, svar. 2 (assay name: Pchry)
Forward Primer PchryF4-1: 5'-GCCTGTCCGAGCGTCACTT
Reverse Primer PchryR8: 5'-CCCCCGGGATCGGAG
Probe PenP6: 5'-CCAACACACAAGCCGTGCTTGAGG
Penicillium chrysogenum/griseofulvum/glandicola/coprophilum/expansum
and Eupenicillium crustaceum/egyptiacum (assay name: PenGrp3)
Forward Primer PchryF1: 5'-CGGGCCCGCCTTAAC
Reverse Primer PchryR1-1: 5'-GAAAGTTTTAAATAATTTATATTTTCACTCAGAGTA
Probe PenP2: 5'-CGCGCCCGCCGAAGACA
Penicillium citreonigrum (Pcteo2)
Forward Primer PcteoF2-1: 5'-GGGGGCATCTGCtCTCTG
Reverse Primer PcteoR2-1: 5'-CCGTTGTTGAAAGTTTTAAGTTATcTAA
Probe* PenAspP1mgb: 5'-CCAACCTCCCACCCGTG
Penicillium citrinum/sartoryi/westlingi (Pcitr)
Forward Primer PcitrF1: 5'-CCGTGTTGCCCGAACCTA
Reverse Primer PcitrR1: 5'-TTGTTGAAAGTTTTAACTAATTTCGTTATAG
Probe PcitrP1: 5'-TGCAACTTCAGACAGCGTTCAGGGG
Penicillium coprophilum (assay name: Pcopr)
Forward Primer PcoprF1-1: 5'-GGGTCCAACCTCCCACtCA
Reverse Primer PchryR1-1: 5'-GAAAGTTTTAAATAATTTATATTTTCACTCAGAgTA
Probe PenP1: 5'-CGCCTTAACTGGCCGCCGG
Penicillium corylophilum (assay name: Pcory)
Forward Primer PcoryF1: 5'-GTCCAACCTCCCACCCA
Reverse Primer PcoryR3-1: 5'-GCTCAGACTGCAATCTTCAGACTGT
Probe PcoryP1: 5'-CTGCCCTCTGGCCCGCG
Penicillium crustosum/camembertii/commune/echinulatum/solitum
(assay name: PenGrp2)
Forward Primer PchryF1: 5'-CGGGCCCGCCTTAAC
Reverse Primer PauraR1-1: 5'-GAAAGTTTTAAATAATTTATATTTTCACTCAGAGTT
Probe PenP2: 5'-CGCGCCCGCCGAAGACA
Penicillium decumbens (assay name: Pdecu3)
Forward Primer PdecuF3: 5'-GGCCTCCGTCCTCCTTTG
Reverse Primer PdecuR4-1: 5'- GACCTACAGAGCGGGTGATGA
Probe PenP7: 5'-CCGAAAGGCAGCGGCGGC
Penicillium digitatum (assay name: Pdigi)
Forward Primer PaethF1-1: 5'-CGGGGGGCTCtCGCT
Reverse Primer PdigiR1: 5'-CGTTGTTGAAAGTTTTAAATAATTTCGT
Probe PenP2: 5'-CGCGCCCGCCGAAGACA
Penicillium expansum (assay name: Pexpa)
Forward Primer PexpaF1-1: 5'-TTACCGAGTGAGGGCCgTT
Reverse Primer PexpaR2-1: 5'-GCCCGCCGAAGCtACG
Probe PenP3: 5'-TCCAACCTCCCACCCGTGTTTATTT
Penicillium fellutanum/charlesii (assay name: Pfell2)
Forward Primer PfellF2: 5'- CTGAGTGCGGGCCCTCT
Reverse Primer PfellR2: 5'- CGCCGAAGCAACACTGTAAG
Probe* PenAspP1mgb: 5'-CCAACCTCCCACCCGTG
Penicillium glandicola (assay name: Pglan)
Forward Primer PglanF1-1: 5'-CCGGGGGGCTTtCGT
Reverse Primer PchryR1: 5'-GAAAGTTTTAAATAATTTATATTTTCACTCAGACTA
Probe PenP2: 5'-CGCGCCCGCCGAAGACA
Penicillium griseofulvum (assay name: Pgris)
Forward Primer PgrisF1-1: 5'-ACCTGCGGAAGGATCATTtCT
Reverse Primer PchryR6: 5'-CCCGGCGGCCAGTT
Probe PenP3: 5'-TCCAACCTCCCACCCGTGTTTATTT
Penicillium implicatum (assay name: Pimpl)
Forward Primer PimplF1: 5'-GCCGAAGACCCCCCTGT
Reverse Primer PimplR1: 5'CGTTGTTGAAAGTTTTGACTGATTGT
Probe PimplP1: 5'-AACGCTGTCTGAAGCTTGCAGTCTGAGC
Penicillium islandicum (assay name: Pisla)
Forward Primer PislaF1: 5'-CGAGTGCGGGTTCGACA
Reverse Primer PislaR1: 5'-GGCAACGCGGTAACGGTAG
Probe PislaP1: 5'-AGCCCAACCTCCCACCCGTG
Penicillium italicum (assay name: Pital)
Forward Primer PitalF1-1: 5'-CTCCCACCCGTGTTTATTTAtCA
Reverse Primer PexpaR1: 5'-TCACTCAGACGACAATCTTCAGG
Probe PenP1: 5'-CGCCTTAACTGGCCGCCGG
Penicillium melinii (assay name: Pmeli)
Forward Primer PmeliF1-1: 5'-CACGGCTTGTGTGTTGGtCT
Reverse Primer PmeliR1: 5'-GGGCCTACAAGAGCGGAA
Probe PenP7: 5'-CCGAAAGGCAGCGGCGGC
Penicillium miczynskii (assay name: Pmicz)
Forward Primer PmiczF1-1: 5'-GTGTTTAACGAACCTTGTTGCaTT
Reverse Primer PmiczR1-1: 5'-CTCAGACTGCATACTTCAGACaGA
Probe PsimpP1: 5'-CCGCCTCACGGCCGCC
Penicillium olsonii (assay name: Polsn)
Forward Primer PolsnF1: 5'-GGCGAGCCTGCCTTCG
Reverse Primer PenR2: 5'-GATCCGTTGTTGAAAGTTTTAAATAATTTATA
Probe PolsnP2: 5'-TCCGCGCTCGCCGGAGAC
Penicillium oxalicum (assay name: Poxal)
Forward Primer PoxalF1: 5'-GGGCCCGCCTCACG
Reverse Primer PoxalR1: 5'-GTTGTTGAAAGTTTTAACTGATTTAGTCAAGTA
Probe PoxalP1: 5'-ACAAGAGTTCGTTTGTGTGTCTTCGGCG
Penicillium purpurogenum (assay name: Ppurp)
Forward Primer PpurpF1: 5'-AGGATCATTACTGAGTGCGGA
Reverse Primer PpurpR1: 5'-GCCAAAGCAACAGGGTATTC
Probe PpurpP1: 5'-CCCTCGCGGGTCCAACCTCC
Penicillium raistrickii (assay name: Prais3)
Forward Primer PgrisF1-1: 5'-ACCTGCGGAAGGATCATTtCT
Reverse Primer PraisR2: 5'-CCGGCGGCCTGACG
Probe PenP3: 5'-TCCAACCTCCCACCCGTGTTTATTT
Penicillium restrictum (assay name: Prest2)
Forward Primer PrestF2-1: 5'-CACGGCTTGTGTGTTGGtCCT
Reverse Primer PrestR1-1: 5'-CGGCCGGGCCTaCAA
Probe PenP7: 5'-CCGAAAGGCAGCGGCGGC
Penicillium roquefortii (assay name: Proqu)
Forward Primer PchryF1: 5'-CGGGCCCGCCTTAAC
Reverse Primer ProquR2: 5'-TTAAATAATTTATATTTGTTCTCAGACTGCAT
Probe PenP2: 5'-CGCGCCCGCCGAAGACA
Penicillium sclerotiorum (assay name: Psclr)
Forward Primer PsclrF1: 5'-TTCCCCCGGGAACAGG
Reverse Primer PsclrR1: 5'-GCCCCATACGCTCGAGGAT
Probe PsclrP1: 5'-CCGAAAGGCAGTGGCGGCAC
Penicillium simplicissimum/ochrochloron (assay name: Psimp2)
Forward Primer PsimpF2-1: 5'-CGCCGAAGACACCATTGAtCT
Reverse Primer PsimpR4-1: 5'-CTGAATTCTGCAATTCACATaACG
Probe PsimpP2: 5'-TGTCTGAAGATTGCAGTCTGAGCGATTAGC
Penicillium glabrum/lividum/pupurescens/spinulosum/thomii (assay
name: Pspin2)
Forward Primer PspinF2: 5'-CATTACTGAGTGAGGGCCCTCT
Reverse Primer PspinR2-1: 5'-CGTGAGGCGGGaGCA
Probe* PenAspP1mgb: 5'-CCAACCTCCCACCCGTG
Penicillium variabile (assay name: Pvarb2)
Forward Primer PvarbF2-1: 5'-TTACCGAGTGCGGGTTCtAA
Reverse Primer PvarbR2: 5'-CGAGGCAACGCGGTAAC
Probe* PenAspP1mgb: 5'-CCAACCTCCCACCCGTG
Penicillium verrucosum, svar. 1 (assay name: Pverr)
Forward Primer PverrF2: 5'-CGGGCCCGCCTTTG
Reverse Primer PauraR1: 5'-GAAAGTTTTAAATAATTTATATTTTCACTCAGACTT
Probe PenP2: 5'-CGCGCCCGCCGAAGACA
Rhizomucor meihei/pusillus/variabilis
(assay name: Rmuc)
Forward Primer NS92F: 5'-CACCGCCCGTCGCTAC
Reverse Primer RmucR1: 5'-GTAGTTTGCCATAGTTCGGCTA
Probe RmucP1: 5'-TTGAATGGCTATAGTGAGCATATGGGAGGCT
Rhizopus stolonifer (assay name: Rstol)
Forward Primer NS92F: 5'-CACCGCCCGTCGCTAC
Reverse Primer RstolR1: 5'-GCTTAGTTTGCCATAGTTCTCTAACAA
Probe MucP1: 5'-CCGATTGAATGGTTATAGTGAGCATATGGGATC
Scopulariopsis asperula (assay name:
SCaspr)
Forward Primer SCbrvF1: 5'-CCCCTGCGTAGTAGATCCTACAT
Reverse Primer SCasprR1: 5'-TCCGAGGTCAAACCATGAGTAA
Probe ScopP1: 5'-TCGCATCGGGTCCCGGCG
Scopulariopsis brevicaulis/fusca (assay name: SCbrv)
Forward Primer SCbrvF1: 5'-CCCCTGCGTAGTAGATCCTACAT
Reverse Primer SCbrvR1: 5'-TCCGAGGTCAAACCATGAAATA
Probe ScopP1: 5'-TCGCATCGGGTCCCGGCG
Scopulariopsis brumptii (assay name: SCbrm)
Forward Primer SCbrmF1: 5'-CCCCTGCGTAGTAGTAAAACCA
Reverse Primer SCbrmR1: 5'-CCGAGGTCAAACATCTTTGG
Probe ScopP1: 5'-TCGCATCGGGTCCCGGCG
Scopulariopsis chartarum (assay name: SCchr)
Forward Primer SCchrF1: 5'-CCCCCTGCGTAGTAGTAAAGC
Reverse Primer SCchrR1: 5'-TCCGAGGTCAAACCATCAAG
Probe ScopP1: 5'-TCGCATCGGGTCCCGGCG
Scopulariopsis sphaerospora (assay name: SCsph)
Forward Primer SCsphF1: 5'-CCCCCTGCGTAGTAGTTTACAA
Reverse Primer SCsphR1: 5'-CCGAGGTCAAACCATCAAAAG
Probe ScopP1: 5'-TCGCATCGGGTCCCGGCG
Stachybotrys chartarum (assay name:
Stac)
Forward Primer StacF4: 5'-TCCCAAACCCTTATGTGAACC
Reverse Primer StacR5: 5'-GTTTGCCACTCAGAGAATACTGAAA
Probe StacP2: 5'-CTGCGCCCGGATCCAGGC
Trichoderma asperellum/hamatum (assay
name: Taspr1)
Forward Primer TasprF1: 5'-CCCAAACCCAATGTGAACGT
Reverse Primer TasprR2-1: 5'-GGACTACAGAAAGAGTTTGGTTGCTT
Probe TridP1: 5'-CCAAACTGTTGCCTCGGCGGG
Trichoderma harzianum (assay name: Tharz)
Forward Primer TharzF1: 5'-TTGCCTCGGCGGGAT
Reverse Primer TharzR1: 5'-ATTTTCGAAACGCCTACGAGA
Probe TharzP1: 5'-CTGCCCCGGGTGCGTCG
Trichoderma longibrachiatum/citrinoviride (assay name: Tlong)
Forward Primer TlongF1: 5'-TGCCTCGGCGGGATTC
Reverse Primer TlongR1: 5'-CGAGAAAGGCTCAGAGCAAAAAT
Probe TlongP1: 5'-TCGCAGCCCCGGATCCCA
Trichoderma viride/atroviride/koningii (assay name: Tviri)
Forward Primer TviriF1: 5'-CCCAAACCCAATGTGAACCA
Reverse Primer TviriR1: 5'-TCCGCGAGGGGACTACAG
Probe TridP1: 5'-CCAAACTGTTGCCTCGGCGGG
Ulocladium atrum (assay name: Uatrm)
Forward Primer UatrmF2: 5'-CGGGCTGGCATCCTTC
Reverse Primer UatrmR2: 5'-CTGATTGCAATTACAAAAGGTTTATG
Probe UloP1: 5'-TGAATTATTCACCCGTGTCTTTTGCGTACTTCT
Ulocladium botrytis (assay name: Ubotr)
Forward Primer UbotrF1: 5'-CCCCCAGCAGTGCGTT
Reverse Primer UbotrR1: 5'-CTGATTGCAATTACAAAAGGTTTATG
Probe UloP1: 5'-TGAATTATTCACCCGTGTCTTTTGCGTACTTCT
Ulocladium chartarum (assay name: Uchar)
Forward Primer UcharF1-1: 5'-AGCGGGCTGGAATCCaTT
Reverse Primer UcharR1-1: 5'-CTGATTGCAATTACAAAAGGTTgAAT
Probe UloP1: 5'-TGAATTATTCACCCGTGTCTTTTGCGTACTTCT
Wallemia sebi (assay name: Wsebi)
Forward Primer WsebiF1: 5'-GGCTTAGTGAATCCTTCGGAG
Reverse Primer WsebiR1: 5'-GTTTACCCAACTTTGCAGTCCA
Probe WsebiP1: 5'-TGTGCCGTTGCCGGCTCAAATAG
Universal Penicillium, Aspergillus and
Paecilomyces varioti (assay name: PenAsp1mgb)
Forward Primer PenAspF1: 5'-CGGAAGGATCATTACTGAGTG
Reverse Primer PenAspR1: 5'-GCCCGCCGAAGCAAC
Probe* PenAspP1mgb: 5'-CCAACCTCCCACCCGTG
* Applied Biosystems MGB probe, labeled with 6FAM reporter and black hole quencher. Suggest labeling of all other probes with 6FAM reporter and TAMRA or black hole quencher.
Suggested probe and primer concentrations for all assays is 80 nM probe, 1000 nM primers. Primers with (5x) designations indicate suggested use of 5x higher concentration.
Lower case letters in primer sequences denote deliberate mismatches with
target sequences.
Haugland, R. A., S. J. Vesper, and L. J. Wymer. 1999. Quantitative measurement of Stachybotrys chartarum conidia using real time detection of PCR products with the TaqMan™ fluorogenic probe system. Molecular and Cellular Probes 13:329-340.
Roe, J., R A. Haugland, S. J. Vesper, and L. J. Wymer. 2001. Quantification of Stachybotrys chartarum conidia in indoor dust using real time, fluorescent probe-based detection of PCR products. Journal of Exposure Analysis and Environmental Epidemiology 11:12-20.
Haugland, R. A., N. E. Brinkman, and S. J. Vesper. 2002. Evaluation of rapid DNA extraction methods for the quantitative detection of fungal cells using real time PCR analysis. Journal of Microbiological Methods 50:319-323.
Brinkman, N. E., R. A. Haugland, L. J. Wymer, M. Byappanahalli, R. L. Whitman, and S. J. Vesper. 2003. Evaluation of a Rapid, Quantitative Real-Time PCR Method for Cellular Enumeration of Pathogenic Candida Species in Water. Applied and Environmental Microbiology 69:1775-1782.
Haugland, R.A., M. Varma, L.J.Wymer and S. Vesper. 2004. Quantitative PCR analysis of selected Aspergillus, Penicillium and Paecilomyces Species. System. Appl. Microbiol. 27:198-210.