title: Development and evaluation of a real-time polymerase chain reaction assay for the rapid detection of Talaromyces marneffei MP1 gene in human plasma.
abstract: Penicilliosis caused by Talaromyces marneffei is a common AIDS-defining illness in South and Southeast Asia. Diagnosis is based on culture which can take up to 14 days for identification, leading to treatment delay and increased mortality. We developed a TaqMan real-time PCR assay targeting the MP1 gene encoding an abundant cell wall protein specific to T. marneffei. The assay's performance was evaluated in MP1-containing plasmids, clinical isolates, and plasma from HIV-infected patients with and without penicilliosis. The assay consistently detected 10 copies of MP1-containing plasmids per reaction and 100 T. marneffei yeast cells per millilitre plasma. There were no amplification with seven other Penicillium species and six other HIV-associated fungal pathogens tested. The assay was evaluated in 70 patients with AIDS: 50 patients with culture-confirmed penicilliosis and 20 patients with opportunistic infections other than penicilliosis. The diagnostic sensitivity was 70.4% (19/27, 95% CI: 51.5-84.1%) and 52.2% (12/23, 95% CI: 33.0-70.8%) in plasma samples collected prior to and within 48 h of antifungal therapy respectively. The diagnostic specificity was 100% (20/20, 95% CI: 83.9-100%). This assay provides a useful tool for the rapid diagnosis of T. marneffei infection and has the potential to improve the management of patients with penicilliosis.
authors.affiliation: Wellcome Trust Major Overseas Program, Oxford University Clinical Research Unit, Ho Chi Minh City, Vietnam.
authors.forename: Ha Thuc Ai
authors.initials: HTA
authors.lastname: Hien
authors.affiliation: Wellcome Trust Major Overseas Program, Oxford University Clinical Research Unit, Ho Chi Minh City, Vietnam.
authors.forename: Tran Tan
authors.initials: TT
authors.lastname: Thanh
authors.affiliation: Wellcome Trust Major Overseas Program, Oxford University Clinical Research Unit, Ho Chi Minh City, Vietnam.
authors.forename: Nguyen Thi Mai
authors.initials: NTM
authors.lastname: Thu
authors.affiliation: Wellcome Trust Major Overseas Program, Oxford University Clinical Research Unit, Ho Chi Minh City, Vietnam.
authors.forename: Ashley
authors.initials: A
authors.lastname: Nguyen
authors.affiliation: Wellcome Trust Major Overseas Program, Oxford University Clinical Research Unit, Ho Chi Minh City, Vietnam.
authors.forename: Nguyen Tat
authors.initials: NT
authors.lastname: Thanh
authors.affiliation: Hospital for Tropical Diseases, Ho Chi Minh City, Vietnam.
authors.forename: Nguyen Phu Huong
authors.initials: NPH
authors.lastname: Lan
authors.affiliation: Wellcome Trust Major Overseas Program, Oxford University Clinical Research Unit, Ho Chi Minh City, Vietnam.|Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia.
authors.forename: Cameron
authors.initials: C
authors.lastname: Simmons
authors.affiliation: Hawaii Center for AIDS, University of Hawaii at Manoa, Honolulu, HI, USA.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Shikuma
authors.affiliation: Hospital for Tropical Diseases, Ho Chi Minh City, Vietnam.
authors.forename: Nguyen Van Vinh
authors.initials: NVV
authors.lastname: Chau
authors.affiliation: Wellcome Trust Major Overseas Program, Oxford University Clinical Research Unit, Ho Chi Minh City, Vietnam.|Centre for Tropical Medicine, Nuffield Department of Clinical Medicine, Oxford University, Oxford, UK.
authors.forename: Guy
authors.initials: G
authors.lastname: Thwaites
authors.affiliation: Wellcome Trust Major Overseas Program, Oxford University Clinical Research Unit, Ho Chi Minh City, Vietnam.|Hawaii Center for AIDS, University of Hawaii at Manoa, Honolulu, HI, USA.|Centre for Tropical Medicine, Nuffield Department of Clinical Medicine, Oxford University, Oxford, UK.
authors.forename: Thuy
authors.initials: T
authors.lastname: Le
mesh_terms: D017088:AIDS-Related Opportunistic Infections / Q000097:blood / Q000175:diagnosis / Q000188:drug therapy / Q000382:microbiology; D000935:Antifungal Agents / Q000627:therapeutic use; D001210:Asia, Southeastern; D005656:Fungal Proteins / Q000097:blood / Q000235:genetics*; D015658:HIV Infections; D006801:Humans; D009181:Mycoses / Q000097:blood / Q000175:diagnosis / Q000188:drug therapy / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D032901:Talaromyces / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D000935:Antifungal Agents; D005656:Fungal Proteins
keywords: 
  HIV
  ; 
  Penicillium marneffei
  ; 
  Talaromyces marneffei
  ; TaqMan real-time polymerase chain reaction