Biomedical subjects
Publications and source records attributed to Kenneth H L Ng.
We describe a case of Solobacterium moorei bacteremia in a 43-year-old woman presenting with acute proctitis complicating radiotherapy for cervical carcinoma. Phenotypic tests failed to identify the bacterium, which was subsequently identified by 16S rRNA gene sequencing. 16S rRNA gene sequencing could help better define the pathogenicity of S. moorei.
Unlike other viridans streptococci, members of the "Streptococcus milleri group" are often associated with abscess formation, but are only rare causes of infective endocarditis. Although it has been shown that almost all S. intermedius isolates and most S. constellatus isolates, but only 19% of S. anginosus isolates, were associated with abscess formation, no report has addressed the relative importance of the 3 species of the "S. milleri group" in infective endocarditis. During a 5-year period (April 1997 through March 2002), 6 cases of "S. milleri" endocarditis (out of 377 cases of infective endocarditis), that fulfil the Duke's criteria for the diagnosis of infective endocarditis, were encountered. All 6 "S. milleri" isolates were identified as S. anginosus by 16S ribosomal RNA (rRNA) gene sequencing. Three patients had underlying chronic rheumatic heart disease and 1 was an IV drug abuser. Five had monomicrobial bacteremia, and 1 had polymicrobial (S. anginosus, S. mitis, Granulicatella adiacens, and Slackia exigua) bacteremia. Two patients died. None of the 6 isolates were identified by the Vitek system (GPI) or the API system (20 STREP) at >95% confidence. All 6 isolates were sensitive to penicillin G (MIC 0.008-0.064 microg/mL), cefalothin, erythromycin, clindamycin, and vancomycin. Accurate identification to the species level, by 16S rRNA gene sequencing, in cases of bacteremia caused by members of the "S. milleri group", would have direct implication on the underlying disease process, hence guiding diagnosis and treatment. Infective endocarditis should be actively looked for in cases of monomicrobial S. anginosus bacteremia, especially if the organism is recovered in multiple blood cultures.
Due to the inadequate automation in the amplification and sequencing procedures, the use of 16S rRNA gene sequence-based methods in clinical microbiology laboratories is largely limited to identification of strains that are difficult to identify by phenotypic methods. In this study, using conventional full-sequence 16S rRNA gene sequencing as the "gold standard," we evaluated the usefulness of the MicroSeq 500 16S ribosomal DNA (rDNA)-based bacterial identification system, which involves amplification and sequencing of the first 527-bp fragment of the 16S rRNA genes of bacterial strains and analysis of the sequences using the database of the system, for identification of clinically significant bacterial isolates with ambiguous biochemical profiles. Among 37 clinically significant bacterial strains that showed ambiguous biochemical profiles, representing 37 nonduplicating aerobic gram-positive and gram-negative, anaerobic, and Mycobacterium species, the MicroSeq 500 16S rDNA-based bacterial identification system was successful in identifying 30 (81.1%) of them. Five (13.5%) isolates were misidentified at the genus level (Granulicatella adiacens was misidentified as Abiotrophia defectiva, Helcococcus kunzii was misidentified as Clostridium hastiforme, Olsenella uli was misidentified as Atopobium rimae, Leptotrichia buccalis was misidentified as Fusobacterium mortiferum, and Bergeyella zoohelcum was misidentified as Rimerella anatipestifer), and two (5.4%) were misidentified at the species level (Actinomyces odontolyticus was misidentified as Actinomyces meyeri and Arcobacter cryaerophilus was misidentified as Arcobacter butzleri). When the same 527-bp DNA sequences of these seven isolates were compared to the known 16S rRNA gene sequences in the GenBank, five yielded the correct identity, with good discrimination between the best and second best match sequences, meaning that the reason for misidentification in these five isolates was due to a lack of the 16S rRNA gene sequences of these bacteria in the database of the MicroSeq 500 16S rDNA-based bacterial identification system. In conclusion, the MicroSeq 500 16S rDNA-based bacterial identification system is useful for identification of most clinically important bacterial strains with ambiguous biochemical profiles, but the database of the MicroSeq 500 16S rDNA-based bacterial identification system has to be expanded in order to encompass the rarely encountered bacterial species and achieve better accuracy in bacterial identification.