title: A comparison of capillary electrophoresis and direct sequencing in upstream conserved sequence region analysis of Pneumocystis jirovecii strains.
abstract: The major surface glycoprotein (MSG) of Pneumocystis jirovecii is the most abundant surface protein and appears to play a critical role in the pathogenesis of pneumocystosis. The expressed MSG gene is located immediately downstream of a region called the upstream conserved sequence (UCS). The UCS contains a region of tandem repeats that vary in number and sequence. In the present study, we have used capillary electrophoresis and direct sequencing to detect the variability in the repeat units of UCS. By direct sequencing the PCR products from samples of 13 patients, we have identified three types of repeat units which consisted of 10 nt and three different patterns in the UCS region with three and four repeats: 1, 2, 3 (84.6 %); 1, 2, 3, 3 (8.2 %); and a new genotype 2, 2, 3, 3 (8.2 %). The same samples were analysed by capillary electrophoresis. Three samples (23 %) contained a mixture of two or three different patterns of UCS repeats. In conclusion, quantifying the number of repeat units in the UCS by capillary electrophoresis provides a potential new method for the rapid typing of P. jirovecii and the detection of mixed infection.
authors: Fungal and Parasitic Molecular Biology Laboratory, School of Medicine, University of Sfax, Magida Boulila Street, 3029 Sfax, Tunisia.
authors: M A
authors: MA
authors: Jarboui
authors: Fungal and Parasitic Molecular Biology Laboratory, School of Medicine, University of Sfax, Magida Boulila Street, 3029 Sfax, Tunisia.
authors: F
authors: F
authors: Mseddi
authors: Fungal and Parasitic Molecular Biology Laboratory, School of Medicine, University of Sfax, Magida Boulila Street, 3029 Sfax, Tunisia.
authors: H
authors: H
authors: Sellami
authors: Fungal and Parasitic Molecular Biology Laboratory, School of Medicine, University of Sfax, Magida Boulila Street, 3029 Sfax, Tunisia.
authors: A
authors: A
authors: Sellami
authors: Laboratory of Immunology, Hedi Chaker hospital, Sfax, Tunisia.
authors: N
authors: N
authors: Mahfoudh
authors: Fungal and Parasitic Molecular Biology Laboratory, School of Medicine, University of Sfax, Magida Boulila Street, 3029 Sfax, Tunisia.
authors: F
authors: F
authors: Makni
authors: Laboratory of Immunology, Hedi Chaker hospital, Sfax, Tunisia.
authors: H
authors: H
authors: Makni
authors: Fungal and Parasitic Molecular Biology Laboratory, School of Medicine, University of Sfax, Magida Boulila Street, 3029 Sfax, Tunisia.
authors: A
authors: A
authors: Ayadi
mesh_terms: D017124:Conserved Sequence; D004271:DNA, Fungal / Q000235:genetics*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005656:Fungal Proteins / Q000235:genetics; D014644:Genetic Variation; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics; D058889:Molecular Typing / Q000379:methods; D016533:Mycological Typing Techniques / Q000379:methods*; D045363:Pneumocystis carinii / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D004271:DNA, Fungal; D005656:Fungal Proteins; D008562:Membrane Glycoproteins