Identification of Mycobacterium neoaurumIsolated from a Neutropenic Patient with Catheter-Related Bacteremia by  16S rRNA Sequencing

Woo, Patrick C. Y.; Tsoi, Hoi-Wah; Leung, Kam Lun; Lum, Peggy N. L.; Leung, Andy S. P.; Ma, Choi-Ha; Kam, Kai‐Man; Yuen, Kwok‐Yung

doi:10.1128/jcm.38.9.3515-3517.2000

Cited by 82 publications

(21 citation statements)

References 24 publications

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“…With the whole genome sequencing data of more than 100 bacteria and archaea available, it has been noted that there are relatively small interoperon heterogeneities for 16S rDNAs. In our experience in 16S rDNA sequencing, both for the known species and novel genera and species, direct sequencing of the PCR products has always been successful (16,(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)30). In this study, when we first amplified the 16S rDNA of HKU15 T and sequenced the PCR product directly, it was repeatedly observed that the 5' end of the PCR product could not be sequenced clearly, with the presence of multiple peaks.…”

Section: Discussionmentioning

confidence: 85%

Anaerospora hongkongensis Gen. Nov. Sp. Nov., a Novel Genus and Species with Ribosomal DNA Operon Heterogeneity Isolated from an Intravenous Drug Abuser with Pseudobacteremia

Woo

Teng

Leung

et al. 2005

Microbiology and Immunology

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Since the discovery of polymerase chain reaction (PCR) and DNA sequencing, comparison of the gene sequences of bacterial species has shown that the 16S rDNA is highly conserved within a species and among species of the same genus, and hence can be used as the new standard for classification of bacteria. Using this new standard, phylogenetic trees, based on base differences between species, are constructed; and bacteria are classified and re-classified into new genera. Recently, the use of this technique for the identification of bacterial strains with ambiguous biochemical profiles and the discovery of novel genera and species have been reported by us and other researchers (5-7, 10, 16, 18-30).Based on the results of 16S rDNA sequencing, major changes in the taxonomy of anaerobic Gram-negative bacteria have been made in recent years (2,4,9,11,15). At the moment, about 50 genera of anaerobic Gram-negative bacteria have been described, three of which were sporulated. In this study, we report the isolation of an anaerobic Gram-negative bacterial strain from the blood culture of an intravenous drug abuser. The strain, named HKU15 T , exhibited phenotypic characteristics that do not fit into patterns of any known genus and species. 16S rDNA sequencing showed that there was only about 90% base identity between the 16S rDNA of HKU15T and that of the most closely related species, Dendrosporobacter quercicolus (15). On the basis of these results we propose a new genus and species, Anaerospora hongkongensis gen. nov. sp. Abstract: A bacterium was isolated from the blood culture of an intravenous drug abuser with pseudobacteremia. The cells were strictly anaerobic, straight or slightly curved, sporulating, Gram-negative rods. It grew on sheep blood agar as non-hemolytic, pinpoint colonies after 48 hr of incubation at 37 C in an anaerobic environment. It was motile but did not produce catalase or cytochrome oxidase. 16S ribosomal DNA (rDNA) sequencing revealed three different copies of 16S rDNA sequences. More than 90% of the differences among them were due to differences in the lengths of the sequences. Phylogenetically, the bacterium is clustered with Dendrosporobacter, Sporomusa, and Propionispora, the other three genera of anaerobic, sporulating, Gram-negative rods. There were 8.6-11.1% differences between the 16S rDNA sequences of the bacterium and that of D. quercicolus, 4.7-15.1% differences between the 16S rDNA sequences of it and those of S. acidovorans, S. aerivorans, S. malonica, S. ovata, S. paucivorans, S. silvacetica, S. spaeroides, and S. termitida, and 7.6-13.1% differences between the 16S rDNA sequences of it and those of P. hippei and P. vibrioides. The G؉C content of the bacterium (mean؎SD) was 46.8؎3.2%. For these reasons, a new genus and species, Anaerospora hongkongensis gen. nov. sp. nov., is proposed, for which HKU15 T is the type strain. Anaerospora hongkongensis

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Section: Discussionmentioning

confidence: 85%

Anaerospora hongkongensis Gen. Nov. Sp. Nov., a Novel Genus and Species with Ribosomal DNA Operon Heterogeneity Isolated from an Intravenous Drug Abuser with Pseudobacteremia

Woo

Teng

Leung

et al. 2005

Microbiology and Immunology

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“…1 Furthermore, non-cultivable organisms and organisms with ambiguous biochemical profiles can be classified and identified. 2 3 Recently, we have reported the use of this technique for the identification of bacterial strains with ambiguous biochemical profiles, [4][5][6][7][8] species that are rarely encountered clinically, [9][10][11][12][13] and a bacterium that is non-cultivable 14 ; the discovery of a novel clinical syndrome 15 16 and two novel species 17 18 ; and the characterisation of β haemolytic Lancefield group G streptococcal bacteraemia 19 and thermotolerant Campylobacter fetus bacteraemia. 20 "The 16S rRNA gene is highly conserved within a species and among species of the same genus, and hence can be used as the new gold standard for speciation of bacteria"…”

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Pseudobacteraemia in a patient with neutropenic fever caused by a novel paenibacillus species: Paenibacillus hongkongensis sp. nov.

Teng

Woo²,

Leung³

et al. 2003

Molecular Pathology

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Aims: To characterise a strain of Gram negative aerobic straight or slightly curved rods (HKU3) isolated from the blood culture of a 9 year old Chinese boy with neutropenic fever and pseudobacteraemia. Methods: The isolate was phenotypically investigated by standard biochemical methods using conventional biochemical tests, scanning electron microscopy, and transmission electron microscopy. Genotypically, the 16S rRNA gene of the bacterium was amplified by the polymerase chain reaction (PCR) and sequenced. The sequence of the PCR product was compared with known 16S rRNA gene sequences in the Genbank by multiple sequence alignment. The G + C content was determined by thermal denaturation. A phylogenetic tree was constructed by the PileUp method.Results: The cells of the bacterial strain were aerobic, sporulating, Gram negative straight or slight curved rods. The bacterium grew on horse blood agar as non-haemolytic, grey colonies of 1 mm in diameter after 24 hours of incubation at 37°C in ambient air. No enhancement of growth was seen in 5% CO 2 . It grew at 50°C as pinpoint colonies after 72 hours of incubation, but did not grow at 65°C or on MacConkey agar. It was non-motile. It produced catalase (weakly positive) and cytochrome oxidase. It reduced nitrate, produced β galactosidase, hydrolysed esculin, and utilised sodium acetate. A scanning electron micrograph of the bacterium showed straight or slightly curved rods. A transmission electron micrograph of the cell wall of the bacterium revealed multiple electron dense layers, including the outer membrane, middle murein layer, and inner cytoplasmic membrane, compatible with its Gram smear appearance. 16S rRNA gene sequencing showed that there were 7.7%, 8.0%, 8.2%, and 8.6% differences between the 16S rRNA gene sequence of the bacterium and those of Paenibacillus macerans, Paenibacillus borealis, Bacillus ehimensis, and Paenibacillus amylolyticus, respectively. The mean (SD) G + C content of the bacterium was 47.6 (2.1) mol%. Phylogenetically, it belongs to the genus paenibacillus (previously called group 3 bacillus). Conclusions: A bacterium that exhibited phenotypic and genotypic characteristics that are very different from closely related members of paenibacillus was the cause of pseudobacteraemia in a patient with neutropenic fever. A new species, Paenibacillus hongkongensis sp. nov. is proposed, for which HKU3 is the type strain.

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“…Pigmented rapidly growing mycobacteria are normally not known to cause disease. However, M. neoaurum, a pigmented rapid grower, has been associated previously with several cases of catheter-related bacteremia and has been best identified using 16S rRNA gene sequencing (25). Definitive identification of these species is difficult without advanced methods such as HPLC or sequencing.…”

Section: Discussionmentioning

confidence: 99%

Phenotypic and Molecular Characterization of Clinical Isolates of Mycobacterium elephantis from Human Specimens

et al. 2002

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Eleven strains of a rapidly growing mycobacterium were isolated from patient specimens originating from various regions of the province of Ontario, Canada, over a 2-year period. Unique high-performance liquid chromatography (HPLC) and PCR-restriction enzyme pattern analysis (PRA) profiles initially suggested a new Mycobacterium species, while sequencing of the 16S rRNA gene revealed a sequence match with Mycobacterium sp. strain MCRO 17 (GenBank accession no. X93028), an isolate determined to be unique which is to date uncharacterized, and also a close similarity to M. elephantis (GenBank accession no. AJ010747), with six base pair variations. A complete biochemical profile of these isolates revealed a species of mycobacteria with phenotypic characteristics similar to those of M. flavescens. HPLC, PRA, and 16S rRNA sequencing of strain M. elephantis DSM 44368T and result comparisons with the clinical isolates revealed that these strains were in fact M. elephantis, a newly described species isolated from an elephant. All strains were isolated from human samples, 10 from sputum and 1 from an axillary lymph node

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Identification of Mycobacterium neoaurumIsolated from a Neutropenic Patient with Catheter-Related Bacteremia by 16S rRNA Sequencing

Cited by 82 publications

References 24 publications

Anaerospora hongkongensis Gen. Nov. Sp. Nov., a Novel Genus and Species with Ribosomal DNA Operon Heterogeneity Isolated from an Intravenous Drug Abuser with Pseudobacteremia

Anaerospora hongkongensis Gen. Nov. Sp. Nov., a Novel Genus and Species with Ribosomal DNA Operon Heterogeneity Isolated from an Intravenous Drug Abuser with Pseudobacteremia

Pseudobacteraemia in a patient with neutropenic fever caused by a novel paenibacillus species: Paenibacillus hongkongensis sp. nov.

Phenotypic and Molecular Characterization of Clinical Isolates of Mycobacterium elephantis from Human Specimens

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