| Clinical relevance of multiple single-nucleotide polymorphisms in Pneumocystis jirovecii Pneumonia: development of a multiplex PCR-single-base-extension methodology. | |
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MedLine Citation:
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PMID: 21389160 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Pneumocystis jirovecii pneumonia (PcP) is a major cause of respiratory illness in patients with AIDS. The identification of multiple single-nucleotide polymorphisms (SNPs) at three distinct P. jirovecii loci encoding dihydrofolate reductase (DHFR), mitochondrial large-subunit rRNA (mtLSU rRNA), and superoxide dismutase (SOD) was achieved using multiplex-PCR (MPCR) followed by direct sequencing and two single-base extension (SBE) techniques. Four SNPs (DHFR312, mt85, SOD215, and SOD110), correlated previously with parameters of disease, were amplified and genotyped simultaneously. The concordance of results between the standard sequencing technique (direct sequencing) and SBE analysis was 96.9% for the acrylamide gel electrophoresis and 98.4% for the capillary electrophoresis. The cross-genetic analysis established several statistical associations among the SNPs studied: mt85C-SOD110T, SOD110T-SOD215C, and SOD110C-SOD215T. These results were confirmed by cluster analysis. Data showed that among the isolates with low to moderate parasite burden, the highest percentages of DHFR312C, mt85C, SOD110T, and SOD215C were detected, whereas for high parasite burden cases the highest frequencies were observed among isolates with DHFR312T, mt85T, SOD110C, and SOD215T. The polymorphisms studied were shown to be suitable genetic targets potentially correlated with PcP clinical data that can be used as predictors of outcome in further studies to help clinical decision-making in the management of PcP. The MPCR/SBE protocol described for the first time in the present study was shown to be a rapid, highly accurate method for genotyping P. jirovecii SNPs encoded by different loci that could be used for epidemiological studies and as an additional procedure for the prognostic classification and diagnosis of PcP. |
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Authors:
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F Esteves; J Gaspar; B De Sousa; F Antunes; K Mansinho; O Matos |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2011-03-09 |
Journal Detail:
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Title: Journal of clinical microbiology Volume: 49 ISSN: 1098-660X ISO Abbreviation: J. Clin. Microbiol. Publication Date: 2011 May |
Date Detail:
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Created Date: 2011-04-28 Completed Date: 2011-07-26 Revised Date: 2011-11-01 |
Medline Journal Info:
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Nlm Unique ID: 7505564 Medline TA: J Clin Microbiol Country: United States |
Other Details:
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Languages: eng Pagination: 1810-5 Citation Subset: IM |
Affiliation:
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Unidade de Protozoários Oportunistas/VIH e Outras Protozooses-CMDT, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, 1349-008 Lisboa, Portugal. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Adult Cluster Analysis DNA, Mitochondrial / genetics DNA, Ribosomal / genetics Female Genotype Humans Male Microbiological Techniques / methods* Middle Aged Molecular Epidemiology / methods Mycological Typing Techniques / methods* Pneumocystis jirovecii / classification*, genetics*, isolation & purification Pneumonia, Pneumocystis / diagnosis, microbiology* Polymerase Chain Reaction / methods* Polymorphism, Single Nucleotide* RNA, Ribosomal, 23S / genetics Superoxide Dismutase / genetics Tetrahydrofolate Dehydrogenase / genetics |
| Chemical | |
Reg. No./Substance:
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0/DNA, Mitochondrial; 0/DNA, Ribosomal; 0/RNA, Ribosomal, 23S; EC 1.15.1.1/Superoxide Dismutase; EC 1.5.1.3/Tetrahydrofolate Dehydrogenase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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