Molecular identification of Penicillium marneffei using rolling circle amplification

TY - JOUR

T1 - Molecular identification of Penicillium marneffei using rolling circle amplification

AU - Sun, J.

AU - Najafzadeh, M.J.

AU - Zhang, J.

AU - Xi, L.

AU - de Hoog, G.S.

N1 - Reporting year: 2010

PY - 2011

Y1 - 2011

N2 - Penicillium marneffei is the aetiological agent of a severe systemic disease in immunocompromised hosts in Southeast Asia. In the present study, we evaluated an identification method based on rolling circle amplification (RCA) enabling rapid and specific detection of single nucleotide differences. Three padlock probes were designed on the basis of the internal transcribed spacers 1 and 2 (ITS) of the rRNA operon. One of these (PmPL1) allowed specific amplification of P. marneffei DNA within one working day using a newly conceived protocol, while no cross-reactivity was observed with other fungi including related biverticillate penicillia. Amplification products can be detected by electrophoresis on agarose gel. The method provides a powerful tool for a rapid specific identification of P. marneffei in the clinical laboratory and has potential for ecological studies. Keywords: Penicillium marneffei; penicilliosis; rapid identification; RCA

AB - Penicillium marneffei is the aetiological agent of a severe systemic disease in immunocompromised hosts in Southeast Asia. In the present study, we evaluated an identification method based on rolling circle amplification (RCA) enabling rapid and specific detection of single nucleotide differences. Three padlock probes were designed on the basis of the internal transcribed spacers 1 and 2 (ITS) of the rRNA operon. One of these (PmPL1) allowed specific amplification of P. marneffei DNA within one working day using a newly conceived protocol, while no cross-reactivity was observed with other fungi including related biverticillate penicillia. Amplification products can be detected by electrophoresis on agarose gel. The method provides a powerful tool for a rapid specific identification of P. marneffei in the clinical laboratory and has potential for ecological studies. Keywords: Penicillium marneffei; penicilliosis; rapid identification; RCA

U2 - 10.1111/j.1439-0507.2011.02017.x

DO - 10.1111/j.1439-0507.2011.02017.x

M3 - Article

SN - 0933-7407

VL - 54

SP - 751

EP - 759

JO - Mycoses

JF - Mycoses

IS - 6

ER -