Specific identification of Penicillium marneffei by a polymerase chain reaction/hybridization technique

N. Vanittanakom, W. G. Merz, N. Sittisombut, C. Khamwan, Kenrad Edwin Nelson, T. Sirisanthana

Research output: Contribution to journalArticle

Abstract

Penicillium marneffei has been described recently as a cause of an emerging mycotic infection in HIV-infected patients. A PCR/hybridization assay was developed to rapidly identify this pathogen. The nucleotide sequence of the 631-bp region of 18S ribosomal DNA of P marneffei was determined using the standard dideoxy chain termination method. An oligonucleotide probe was designed on the basis of the analysed sequences of P marneffei and 18S rDNA sequences of other fungi in the GenBank database. A 631-bp PCR product was amplified using primers RRF1 and RRH1 from P marneffei and seven other fungi, Penicillium spp., Aspergillus fumigatus, A. flavus, Histoplasma capsulatum, Cryptococcus neoformans, Candida albicans and C. krusei. A 15 oligonucleotide segment (Pm3) which was specific for P marneffei was synthesized and used as a probe. Only the PCR products of P marneffei isolates hybridized with the Pm3 oligonucteotide probe. The sensitivity of the assay was approximately 0.5 pg/μl and 0.1 pg/μl of DNA by PCR and Southern hybridization, respectively. The usefulness of this method as a diagnostic tool will require further studies.

Original languageEnglish (US)
Pages (from-to)169-175
Number of pages7
JournalMedical Mycology
Volume36
Issue number3
DOIs
StatePublished - Jun 1998

Fingerprint

Penicillium
hybridization
polymerase chain reaction
Candida krusei
Histoplasma capsulatum
nucleotide sequences
Polymerase Chain Reaction
fungi
oligonucleotide probes
Cryptococcus neoformans
Aspergillus fumigatus
HIV infections
assays
Aspergillus flavus
oligonucleotides
Ribosomal DNA
Candida albicans
ribosomal DNA
Southern blotting
Fungi

Keywords

  • Hybridization technique
  • Penicillium marneffei
  • Polymerase chain reaction

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • veterinary(all)
  • Microbiology

Cite this

Specific identification of Penicillium marneffei by a polymerase chain reaction/hybridization technique. / Vanittanakom, N.; Merz, W. G.; Sittisombut, N.; Khamwan, C.; Nelson, Kenrad Edwin; Sirisanthana, T.

In: Medical Mycology, Vol. 36, No. 3, 06.1998, p. 169-175.

Research output: Contribution to journalArticle

Vanittanakom, N. ; Merz, W. G. ; Sittisombut, N. ; Khamwan, C. ; Nelson, Kenrad Edwin ; Sirisanthana, T. / Specific identification of Penicillium marneffei by a polymerase chain reaction/hybridization technique. In: Medical Mycology. 1998 ; Vol. 36, No. 3. pp. 169-175.
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