Mycobacterium neoaurum is rapidly growing mycobacteria that can cause human infections. It commonly causes bloodstream infections in immunocompromised hosts, and unlike other mycobacteria species, it rarely causes pulmonary infections. We confirmed the first pulmonary infection case in Korea caused by M. neoaurum using full-length 16S rRNA gene sequencing.
Adult ; Female ; Humans ; Lung Diseases/*diagnosis/microbiology ; Mycobacterium/genetics/*isolation & purification ; Mycobacterium Infections/*diagnosis/microbiology ; Nontuberculous Mycobacteria/genetics/isolation & purification ; RNA, Ribosomal, 16S/genetics ; Republic of Korea ; Sequence Analysis, RNA

OBJECTIVETo explore the causative pathogens in littoral hand infections which exhibited chronic granulomatous inflammation, the relationship between chronic granulomatous inflammation and mycobacteria and to discuss the prospects of PCR in clinical application for diagnosis of granulomatous inflammation.

METHODWith 16S-rDNA as the target sequence, Nest-PCR was used to detect mycobacteria directly from 37 cases of chronic granulomatous inflammations, and identified them by gene sequencing.

RESULTSTwenty-four of 37 cases were positive for mycobacteria by Nest-PCR, in which 17 were M.marinum, 1 M.chelonae, 2 M.avium, 2 M.kansasii, and 2 M.tubercular through gene sequencing.

CONCLUSIONSNest-PCR combining gene sequencing proved to be a liable and sensitive method to detect Non-tubercular mycobacteria (NTM) in fresh tissue. NTM is the major factor of hand specific chronic infections other than tubercular. Pathological changes are difficult to differentiate TB from NTM and bacterial evidence was necessary.

Chronic Disease ; DNA, Bacterial ; chemistry ; genetics ; Granuloma ; diagnosis ; microbiology ; Hand ; Humans ; Inflammation ; diagnosis ; microbiology ; Molecular Diagnostic Techniques ; Mycobacterium Infections, Nontuberculous ; diagnosis ; microbiology ; Mycobacterium marinum ; genetics ; isolation & purification ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Nontuberculous Mycobacteria ; genetics ; isolation & purification ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; genetics ; Sequence Analysis, DNA

Mycobacterium longobardum is a slow-growing, nontuberculous mycobacterium that was first characterized from the M. terrae complex in 2012. We report a case of M. longobardum induced chronic osteomyelitis. A 71-yr-old man presented with inflammation in the left elbow and he underwent a surgery under the suspicion of tuberculous osteomyelitis. The pathologic tissue culture grew M. longobardum which was identified by analysis of the 65-kDa heat shock protein and full-length 16S rRNA genes. The patient was cured with the medication of clarithromycin and ethambutol without further complications. To the best of our knowledge, this is the first report of a M. longobardum infection worldwide.
Aged ; Anti-Bacterial Agents/therapeutic use ; Bacterial Proteins/genetics ; Chaperonin 60/genetics ; Clarithromycin/therapeutic use ; Elbow/pathology ; Ethambutol/therapeutic use ; Humans ; Male ; Mycobacterium Infections, Nontuberculous/*microbiology ; Nontuberculous Mycobacteria/classification/genetics/*isolation & purification ; Osteomyelitis/diagnosis/drug therapy/*microbiology/pathology ; RNA, Ribosomal, 16S/genetics ; Treatment Outcome

BACKGROUND: A peptide nucleic acid (PNA) probe-based real-time PCR (PNAqPCR(TM) TB/NTM detection kit; PANAGENE, Korea) assay has been recently developed for the simultaneous detection of Mycobacterium tuberculosis complex (MTBC) and nontuberculous mycobacteria (NTM) in clinical specimens. The study was aimed at evaluation of the performance of PNA probe-based real-time PCR in respiratory specimens. METHODS: To evaluate potential cross-reactivity, the extracted DNA specimens from Mycobacterium species and non-mycobacterial species were tested using PNA probe-based real-time PCR assay. A total of 531 respiratory specimens (482 sputum specimens and 49 bronchoalveolar washing fluid specimens) were collected from 230 patients in July and August, 2011. All specimens were analyzed for the detection of mycobacteria by direct smear examination, mycobacterial culture, and PNA probe-based real-time PCR assay. RESULTS: In cross-reactivity tests, no false-positive or false-negative results were evident. When the culture method was used as the gold standard test for comparison, PNA probe-based real-time PCR assay for detection of MTBC had a sensitivity and specificity of 96.7% (58/60) and 99.6% (469/471), respectively. Assuming the combination of culture and clinical diagnosis as the standard, the sensitivity and specificity of the new real-time PCR assay for detection of MTBC were 90.6% (58/64) and 99.6% (465/467), respectively. The new real-time PCR for the detection of NTM had a sensitivity and specificity of 69.0% (29/42) and 100% (489/489), respectively. CONCLUSIONS: The new real-time PCR assay may be useful for the detection of MTBC in respiratory specimens and for discrimination of NTM from MTBC.
Bronchoalveolar Lavage Fluid/microbiology ; DNA Probes/chemistry/metabolism ; DNA, Bacterial/*analysis ; Humans ; Molecular Typing/*methods ; Mycobacterium tuberculosis/*genetics/isolation & purification ; Nontuberculous Mycobacteria/*genetics/isolation & purification ; Nucleic Acid Hybridization ; Peptide Nucleic Acids/chemistry/*metabolism ; *Real-Time Polymerase Chain Reaction ; Respiratory System/*microbiology ; Sputum/microbiology

Objective: To understand the etiological characteristics and drug susceptibility of Mycobacterium thermoresistibile and Mycobacterium elephantis isolated from a cow with mastitis and provide evidence for the prevention and control of infectious mastitis in cows. Methods: The milk sample was collected from a cow with mastitis, which was pretreated with 4% NaOH and inoculated with L-J medium for Mycobacterium isolation. The positive cultures were initially identified by acid-fast staining and multi-loci PCR, then Mycobacterium species was identified by the multiple loci sequence analysis (MLSA) with 16S rRNA, hsp65, ITS and SodA genes. The drug sensitivity of the isolates to 27 antibiotics was tested by alamar blue assay. Results: Two anti-acid stain positive strains were isolated from the milk of a cow with mastitis, which were identified as non-tuberculosis mycobacterium by multi-loci PCR, and multi-loci nucleic acid sequence analysis indicated that one strain was Mycobacterium thermoresistibile and another one was Mycobacterium elephantis. The results of the drug susceptibility test showed that the two strains were resistant to most antibiotics, including rifampicin and isoniazid, but they were sensitive to amikacin, moxifloxacin, levofloxacin, ethambutol, streptomycin, tobramycin, ciprofloxacin and linezolid. Conclusions:Mycobacterium thermoresistibile and Mycobacterium elephantis were isolated in a cow with mastitis and the drug susceptibility spectrum of the pathogens were unique. The results of the study can be used as reference for the prevention and control the infection in cows.
Animals ; Anti-Bacterial Agents/pharmacology* ; Antitubercular Agents/pharmacology* ; Cattle ; Drug Resistance, Bacterial ; Female ; Humans ; Mastitis, Bovine/microbiology* ; Microbial Sensitivity Tests ; Milk/microbiology* ; Mycobacterium/isolation & purification* ; Mycobacterium Infections/veterinary* ; Mycobacterium tuberculosis/drug effects* ; Nontuberculous Mycobacteria/isolation & purification* ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics*