Development and evaluation of a real-time polymerase chain reaction assay for the rapid detection of Talaromyces marneffei MP1 gene in human plasma

Mycoses. 2016 Dec;59(12):773-780. doi: 10.1111/myc.12530. Epub 2016 Jul 25.

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.

Keywords: HIV; Penicillium marneffei; Talaromyces marneffei; TaqMan real-time polymerase chain reaction.

Publication types

  • Evaluation Study

MeSH terms

  • AIDS-Related Opportunistic Infections / blood
  • AIDS-Related Opportunistic Infections / diagnosis
  • AIDS-Related Opportunistic Infections / drug therapy
  • AIDS-Related Opportunistic Infections / microbiology
  • Antifungal Agents / therapeutic use
  • Asia, Southeastern
  • Fungal Proteins / blood
  • Fungal Proteins / genetics*
  • HIV Infections
  • Humans
  • Mycoses / blood
  • Mycoses / diagnosis
  • Mycoses / drug therapy
  • Mycoses / microbiology*
  • Real-Time Polymerase Chain Reaction / methods*
  • Talaromyces / classification
  • Talaromyces / genetics
  • Talaromyces / isolation & purification*

Substances

  • Antifungal Agents
  • Fungal Proteins

Associated data

  • GENBANK/DQ08822
  • GENBANK/DQ08845