Design of a molecular method for subspecies specific identification of Klebsiella pneumoniae by using the 16S ribosomal subunit gene
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| Format: | Artículo científico |
| Language: | en |
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Universidad del Valle
2009
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| _version_ | 1876458867568672768 |
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| author | NELSON ENRIQUE ARENAS |
| author_facet | NELSON ENRIQUE ARENAS |
| contents | Design of a molecular method for subspecies specific identification of Klebsiella pneumoniae by using the 16S ribosomal subunit gene NELSON ENRIQUE ARENAS JUAN CARLOS POLANCO SANDRA MILENA CORONADO CLARA JULIANA DURANGO ARLEY GÓMEZ Medicina Diagnosis pneumoniae ozaenae pneumoniae pneumoniae Klebsiella pneumoniae rhinoscleromatis Introduction: Rhinoscleroma is caused by Klebsiella pneumoniae rhinoscleromatis and the ozena infections caused by K. pneumoniae ozaenae, both infections affect the upper respiratory tract. In the first clinical phases the symptoms are unspecific, and the disease can be misdiagnosed as a common cold, therefore antimicrobial therapy cannot reach effectiveresults and patients must be following up for several years since the infection became chronic.Objective: To identify Klebsiella subspecies using a specific assay based on amplicons restrictionof a gene which encodes 16S subunit ribosomal (rDNA16S).Methodology: Specific restriction patterns were generated; using reported sequences from rDNA16S gene and bioinformatics programs MACAW, PFE, GENEDOC and GENE RUNNER. Amplification and restriction assays were standardized.Results: Predictions in silico allowed to propose an algorithm for Klebsiella species and subspecies identification. Two reference strains were included and two clinical isolates which were biotyped and identified by the proposed method. rDNA16S gene restriction patterns showed differences regarding the initially identified species for conventional methods. Additionally two patterns of bands were observed for K. pneumoniae rhinoscleromatis, indicating the polymorphisms presence n the rDNA16S gene.Conclusions: It was confirmed the difficulty to identify K. pneumoniae subspecies by conventional methods. Implementation of this technique could allow an accurate and rapid differentiation among K. pneumoniae ozaenae and K. pneumoniae rhinoscleromatis aetiological agents of two frequently misdiagnosed infections. Antimicrobial therapy usually could be ineffective, especially in chronic patients. Finally it is considered very important to enlarge the study by using more clinical and reference strains. 2009 artículo científico 0120-8322 https://www.redalyc.org/articulo.oa?id=28340207 en http://www.redalyc.org/revista.oa?id=283 Colombia Médica application/pdf Universidad del Valle Colombia Médica (Colombia) Num.2 Vol.40 |
| format | Artículo científico |
| id | redalyc_28340207 |
| institution | Redalyc |
| language | en |
| publishDate | 2009 |
| publisher | Universidad del Valle |
| spellingShingle | Design of a molecular method for subspecies specific identification of Klebsiella pneumoniae by using the 16S ribosomal subunit gene NELSON ENRIQUE ARENAS Medicina Diagnosis pneumoniae ozaenae pneumoniae pneumoniae Klebsiella pneumoniae rhinoscleromatis Design of a molecular method for subspecies specific identification of Klebsiella pneumoniae by using the 16S ribosomal subunit gene NELSON ENRIQUE ARENAS JUAN CARLOS POLANCO SANDRA MILENA CORONADO CLARA JULIANA DURANGO ARLEY GÓMEZ Medicina Diagnosis pneumoniae ozaenae pneumoniae pneumoniae Klebsiella pneumoniae rhinoscleromatis Introduction: Rhinoscleroma is caused by Klebsiella pneumoniae rhinoscleromatis and the ozena infections caused by K. pneumoniae ozaenae, both infections affect the upper respiratory tract. In the first clinical phases the symptoms are unspecific, and the disease can be misdiagnosed as a common cold, therefore antimicrobial therapy cannot reach effectiveresults and patients must be following up for several years since the infection became chronic.Objective: To identify Klebsiella subspecies using a specific assay based on amplicons restrictionof a gene which encodes 16S subunit ribosomal (rDNA16S).Methodology: Specific restriction patterns were generated; using reported sequences from rDNA16S gene and bioinformatics programs MACAW, PFE, GENEDOC and GENE RUNNER. Amplification and restriction assays were standardized.Results: Predictions in silico allowed to propose an algorithm for Klebsiella species and subspecies identification. Two reference strains were included and two clinical isolates which were biotyped and identified by the proposed method. rDNA16S gene restriction patterns showed differences regarding the initially identified species for conventional methods. Additionally two patterns of bands were observed for K. pneumoniae rhinoscleromatis, indicating the polymorphisms presence n the rDNA16S gene.Conclusions: It was confirmed the difficulty to identify K. pneumoniae subspecies by conventional methods. Implementation of this technique could allow an accurate and rapid differentiation among K. pneumoniae ozaenae and K. pneumoniae rhinoscleromatis aetiological agents of two frequently misdiagnosed infections. Antimicrobial therapy usually could be ineffective, especially in chronic patients. Finally it is considered very important to enlarge the study by using more clinical and reference strains. 2009 artículo científico 0120-8322 https://www.redalyc.org/articulo.oa?id=28340207 en http://www.redalyc.org/revista.oa?id=283 Colombia Médica application/pdf Universidad del Valle Colombia Médica (Colombia) Num.2 Vol.40 |
| title | Design of a molecular method for subspecies specific identification of Klebsiella pneumoniae by using the 16S ribosomal subunit gene |
| topic | Medicina Diagnosis pneumoniae ozaenae pneumoniae pneumoniae Klebsiella pneumoniae rhinoscleromatis |
| url | https://www.redalyc.org/articulo.oa?id=28340207 |